Paul R Langlais

Interests

Research

The role of insulin is to lower blood glucose levels by stimulating glucose uptake into muscle and adipose tissue. Resistance to insulin, a phenomenon directly involved in the pathogenesis of type 2 diabetes, remains to be understood. Basic research has yet to fully discover how insulin action is elicited. Research in the laboratory of Paul R. Langlais, Ph.D., focuses on the identification and characterization of proteins involved in insulin signal transduction and also tests whether the dysfunction of these proteins is involved in the pathogenesis of insulin resistance and type 2 diabetes.Dr. Langlais specializes in the use of mass spectrometry to perform proteomics, a technique that allows for large-scale quantitative analysis of protein abundances between different treatments. This approach led him to the discovery that CLIP-associating protein 2 (CLASP2) is responsive to insulin stimulation, and his now-published findings support the involvement of CLASP2 in insulin-stimulated glucose uptake. Current research is aimed at discovering the role of CLASP2 in insulin action, in addition to identifying new proteins previously unknown to function in this system.Dr. Langlais leads the University of Arizona Department of Medicine Quantitative Proteomics Laboratory, a collaborative environment for investigators at the University of Arizona and their colleagues to perform large-scale quantitative proteomics studies.

Teaching

Insulin action, post-translational modification, signal transduction, cytoskeletal dynamics, glucose uptake, mass spectrometry, quantitative proteomics

Courses

Scholarly Contributions

Chapters

• Langlais, P. R., & Mandarino, L. J. (2019). Proteomic Profiling of Human Skeletal Muscle in Health and Disease. In Cell Omics approaches to understanding muscle biology.

Journals/Publications

• Ahmed, T., Ramonett, A., Kwak, E., Cruz-Flores, P., Ortiz, H. R., Langlais, P. R., Hund, T. J., Mythreye, K., & Lee, N. Y. (2023). Endothelial tip/stalk cell selection requires BMP9-induced β_IV-spectrin expression during sprouting angiogenesis. Molecular Biology of the Cell.
• Barakati, N., Zapata Bustos, R., Coletta, D. K., Langlais, P. R., Kohler, L. N., Luo, M., Funk, J. L., Willis, W. T., & Mandarino, L. J. (2023). Fuel Selection in Skeletal Muscle Exercising at Low Intensity; Reliance on Carbohydrate in Very Sedentary Individuals. Metabolic Syndrome and Related Disorders.
• Iannuzo, N., Dy, A. B., Guerra, S., Langlais, P. R., & Ledford, J. (2023). The Impact of CC16 on Pulmonary Epithelial-Driven Host Responses during Mycoplasma Pneumoniae Infection in Mouse Tracheal Epithelial Cells. Cells.
• Jensen, C., Clements, A. N., Liou, H., Ball, L. E., Bethard, J. R., Langlais, P. R., Toth, R. K., Chauhan, S. S., Casillas, A. L., Daulat, S. R., Kraft, A., Cress, A. E., Miranti, C., Mouneimne, G., Rogers, G. C., & Warfel, N. A. (2023). PIM kinases regulate actin dynamics and tumor cell invasion in hypoxia. Journal of Cell Biology.
• Kwak, E., Ahmed, T., Cruz-Flores, P., Ortiz, H. R., Langlais, P. R., & Lee, N. Y. (2023). Beta IV spectrin inhibits the metastatic growth of melanoma by suppressing VEGFR2-driven tumor angiogenesis. Cancer Med.
• Ledford, J., Langlais, P. R., Cusanovich, D. A., Li, X., Guerra, S., Johnson, M. D., Li, N., Welfley, H., & Iannuzo, N. (2023). CC16 drives VLA-2-dependent SPLUNC1 expression. Frontiers in Immunology.
• Levine, A. A., Liktor-Busa, E., Balasubramanian, S., Palomino, S. M., Burtman, A. M., Couture, S., Lipinski, A. A., Langlais, P. R., & Milnes, T. M. (2023). Depletion of Endothelial-Derived 2-AG Reduces Blood-Endothelial Barrier Integrity via Alteration of VE-Cadherin and the Phospho-Proteome. Int J Mol Sci.
• Mouneimne, G., Langlais, P. R., Wolgemuth, C. W., Padi, M., Roman, M. R., Parker, J. D., Wang, A., Grant, A. D., Ly, K. T., & Parker, S. S. (2021). EVL and MIM/MTSS1 regulate actin cytoskeletal remodeling to promote dendritic filopodia in developing neurons. Journal of Cell Biology.
• Ou, J., Lewandowski, E., Hu, Y., Lipinski, A., Aljasser, A., Colon-Ascanio, M., Morgan, R., Jacobs, L., Zhang, X., Bikowitz, M., Langlais, P. R., Tan, H., Wang, J., Chen, Y., & Choy, J. S. (2023). A yeast-based system to study SARS-CoV-2 Mpro structure and to identify nirmatrelvir resistant mutaations. PLoS Pathogens.
• Parker, S. S., Ly, K. T., Grant, A. D., Wang, A., Parker, J. D., Roman, M. R., Padi, M., Wolgemuth, C. W., Langlais, P. R., & Mouneimne, G. (2023). EVL and MIM/MTSS1 regulate actin cytoskeletal remodeling to promote dendritic filopodia in developing neurons. Journal of Cell Biology.
• Sangam, s., Sun, X., Schwantes-An, T. H., Yegambaram, M., Lu, Q., Zagorski, J., Morrisroe, S. A., Shi, Y., Cook, T., Fisher, A., Frump, A. L., Coleman, A., Sun, Y., Liang, S., Crawford, H., Lutz, K. A., Muan, A. D., Pauciulo, M. W., Karnes, J. H., , Chaudhary, K. R., et al. (2023). SOX17 Deficiency Mediates Pulmonary Hypertension: At the Crossroads of Sex, Metabolism, and Genetics. Am J Respir Crit Care Med.
• Skaria, R. S., Lopez-Pier, M. A., Kathuria, B. S., Leber, C. J., Langlais, P. R., Aras, S. G., Khalpey, Z. I., Hitscherich, P. G., Chnari, E., Long, M., Churko, J., Runyan, R. B., & Konhilas, J. P. (2023). Epicardial Placement of Human Placental Membrane Protects from Heart Injury in a Swine Model of Myocardial Infarction. Physiol Rep.
• Zapata Bustos, R., Coletta, D. K., Galons, J., Davidson, L., Langlais, P. R., Funk, J. L., Willis, W. T., & Mandarino, L. J. (2023). Nonequilibrium thermodynamics and mitochondrial protein content predict insulin sensitivity and fuel selection during exercise in human skeletal muscle. Front Physiol.
• Ahmed, T., Ahmed, T., Cruz-Flores, P., Cruz-Flores, P., Pan, C. C., Pan, C. C., Ortiz, H. R., Ortiz, H. R., Lee, Y. S., Lee, Y. S., Langlais, P. R., Langlais, P. R., Mythreye, K., Mythreye, K., Lee, N. Y., & Lee, N. Y. (2022). EPDR1 is a noncanonical effector of insulin-mediated angiogenesis regulated by an endothelial-specific TGF-β receptor complex.. Journal of Biological Chemistry.
• Cornejo, N. R., Amofah, B., Lipinksi, A. A., Langlais, P. R., Ghosh, I., & Jewett, J. C. (2022). Direct intracellular delivery of benzene diazonium ions as observed by increased tyrosine phosphorylation. Biochemistry.
• Keresztes, A., Olson, K., Nguyen, P., Lopez-Pier, M. A., Hecksel, R., Barker, N. K., Liu, Z., Hruby, V., Konhilas, J. P., Langlais, P. R., & Streicher, J. M. (2022). Antagonism of the Mu-Delta Opioid Receptor Heterodimer Enhances Opioid Anti-Nociception by Activating Src and CaMKII Signaling. Pain.
• Kwak, E. A., Pan, C. C., Ramonett, A., Kumar, S., Cruz-Flores, P., Ahmed, T., Ortiz, H. R., Lochhead, J. L., Ellis, N., Mouneimne, G., Lee, Y. S., Vanderah, T. W., Milnes, T. M., Mohler, P. J., Hund, T. J., Langlais, P. R., Mythreye, K., & Lee, N. Y. (2022). BIV-spectrin as a stalk cell-intrinsic regulator of VEGF signaling. Nature Communications.
• Ramonett, A., Kwak, E. A., Ahmed, T., Flores, P. C., Ortiz, H. R., Lee, Y. S., Vanderah, T. W., Milnes, T. M., Kashatus, D. F., Langlais, P. R., Mythreye, K., & Lee, N. Y. (2022). Regulation of mitochondrial fission by GIPC-mediated Drp1 retrograde transport. Molecular biology of the cell, 33(1), ar4.
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  Dynamin-related protein 1 (Drp1) is a key regulator of mitochondrial fission, a large cytoplasmic GTPase recruited to the mitochondrial surface via transmembrane adaptors to initiate scission. While Brownian motion likely accounts for the local interactions between Drp1 and the mitochondrial adaptors, how this essential enzyme is targeted from more distal regions like the cell periphery remains unknown. Based on proteomic interactome screening and cell-based studies, we report that GAIP/RGS19-interacting protein (GIPC) mediates the actin-based retrograde transport of Drp1 toward the perinuclear mitochondria to enhance fission. Drp1 interacts with GIPC through its atypical C-terminal PDZ-binding motif. Loss of this interaction abrogates Drp1 retrograde transport resulting in cytoplasmic mislocalization and reduced fission despite retaining normal intrinsic GTPase activity. Functionally, we demonstrate that GIPC potentiates the Drp1-driven proliferative and migratory capacity in cancer cells. Together, these findings establish a direct molecular link between altered GIPC expression and Drp1 function in cancer progression and metabolic disorders.
• Riegel, A. C., Vanderah, T. W., Milnes, T. M., Langlais, P. R., Smith, A., Majuta, L., Franca-Solomon, G., Barber, K. R., & Vizcarra, V. S. (2022). Targeting 5-HT2A receptors and Kv7 channels in PFC to attenuate chronic neuropathic pain in rats using a spared nerve injury model.. Neuroscience Letters.
• Vizcarra, V. S., Barber, K. R., Franca-Solomon, G., Majuta, L., Smith, A., Langlais, P. R., Milnes, T. M., Vanderah, T. W., & Riegel, A. C. (2022). Targeting 5-HT2A receptors and Kv7 channels in PFC to attenuate chronic neuropathic pain in rats using a spared nerve injury model. . Neuroscience Letters.
• Wahl, J. R., Vivek, A., Palomino, S. M., Almuslim, M., Cottier, K. E., Langlais, P. R., Streicher, J. M., Vanderah, T. W., & Milnes, T. M. (2022). Extracellular Alterations in pH and K+ Modify the Murine Brain Endothelial Cell Total and Phospho-Proteome. Biology of sex differences.
• Batty, S. R., & Langlais, P. R. (2021). Microtubules in insulin action: what's on the tube?. Trends in endocrinology and metabolism: TEM, 32(10), 776-789.
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  Microtubules (MT) have a role in the intracellular response to insulin stimulation and subsequent glucose transport by glucose transporter 4 (GLUT4), which resides in specialized storage vesicles that travel through the cell. Before GLUT4 is inserted into the plasma membrane for glucose transport, it undergoes complex trafficking through the cell via the integration of cytoskeletal networks. In this review, we highlight the importance of MT elements in insulin action in adipocytes through a summary of MT depolymerization studies, MT-based GLUT4 movement, molecular motor proteins involved in GLUT4 trafficking, as well as MT-related phenomena in response to insulin and links between insulin action and MT-associated proteins.
• Blawn, K. T., Kellohen, K. L., Galloway, E. A., Verkhovsky, V. G., Wahl, J., Vivek, A., Barker, N. K., Cottier, K. E., Vallecillo, T. G., Langlais, P. R., Liktor-Busa, E., Vanderah, T. W., & Milnes, T. M. (2021). Sex hormones regulate NHE1 functional expression and brain endothelial proteome to control paracellular integrity of the blood endothelial barrier. Biology of Sex Differences.
• Casillas, A. L., Chauhan, S. S., Toth, R. K., Sainz, A. G., Clements, A. N., Jensen, C. C., Langlais, P. R., Miranti, C. K., Cress, A. E., & Warfel, N. A. (2021). Direct phosphorylation and stabilization of HIF-1a by PIM1 kinase drives angiogenesis in solid tumors. Oncogene.
• Dy, A. B., Langlais, P. R., Barker, N. K., Addison, K. J., Tanyaratsrisakul, S., Boitano, S. A., Christenson, S. A., Kraft, M., Meyers, D., Bleecker, E., Li, X., & Ledford, J. (2021). Myeloid-associated differentiation marker is a novel SP-A-associated transmembrane protein whose expression on airway epithelial cells correlates with asthma severity. Scientific reports, 11(1), 23392.
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  Surfactant protein A (SP-A) is well-known for its protective role in pulmonary immunity. Previous studies from our group have shown that SP-A mediates eosinophil activities, including degranulation and apoptosis. In order to identify potential binding partners on eosinophils for SP-A, eosinophil lysates were subjected to SP-A pull-down and tandem mass spectrometry (MS/MS) analysis. We identified one membrane-bound protein, myeloid-associated differentiation marker (MYADM), as a candidate SP-A binding partner. Blocking MYADM on mouse and human eosinophils ex vivo prevented SP-A from inducing apoptosis; blocking MYADM in vivo led to increased persistence of eosinophilia and airway hyper-responsiveness in an ovalbumin (OVA) allergy model and increased airways resistance and mucus production in a house dust mite (HDM) asthma model. Examination of a subset of participants in the Severe Asthma Research Program (SARP) cohort revealed a significant association between epithelial expression of MYADM in asthma patients and parameters of airway inflammation, including: peripheral blood eosinophilia, exhaled nitric oxide (FeNO) and the number of exacerbations in the past 12 months. Taken together, our studies provide the first evidence of MYADM as a novel SP-A-associated protein that is necessary for SP-A to induce eosinophil apoptosis and we bring to light the potential importance of this previously unrecognized transmembrane protein in patients with asthma.
• Finlayson, J., Barakati, N., Langlais, P. R., Funk, J. L., Zapata Bustos, R., Coletta, D. K., Luo, M., Willis, W. T., & Mandarino, L. J. (2021). Site-specific acetylation of adenine nucleotide translocase 1 at lysine 23 in human muscle. Analytical biochemistry, 630, 114319.
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  Evidence suggests acetylation of human adenine nucleotide translocase 1 (ANT1) at lysine 23 (Lys23) reduces binding of ADP. Lys23 contributes to the positive charge that facilitates this interaction. This study was undertaken to characterize ANT1 abundance and acetylation by a novel method using small amounts of human skeletal muscle biopsies. Lysates of whole muscle or mitochondria from the same tissue were prepared from needle biopsies of vastus lateralis muscle of healthy volunteers. Lysed proteins were resolved on gels, the section containing ANT1 (surrounding 30 Kd) was excised, digested with trypsin, spiked with labeled unacetylated and acetylated synthetic standard peptides and analyzed by mass spectrometry. Natural logarithm transformation of data linearized ion intensities over a 10-fold range of peptide mass. Coefficients of variation ranged from 7 to 30% for ANT1 abundance and Lys23 acetylation. In three volunteers, ANT1 content was 8.36 ± 0.33 nmol/g wet weight muscle and 0.64 ± 0.05 nmol/mg mitochondria, so mitochondrial content was 13.3 ± 2.4 mg mitochondria per gram muscle. Acetylation of Lys23 averaged 14.3 ± 4.2% and 4.87 ± 1.84% in whole muscle and mitochondria, respectively. This assay makes it possible to assess effects of acetylation on the function of ANT1 in human muscle.
• He, S., Ryu, J., Liu, J., Luo, H., Lv, Y., Langlais, P. R., Wen, J., Dong, F., Sun, Z., Xia, W., Lynch, J. L., Duggirala, R., Nicolson, B., Zhang, M., Shi, Y., Zhang, F., Liu, F., Bai, J., & Dong, L. Q. (2021). LRG1 is an adipokine that mediates obesity-induced hepatosteatosis and insulin resistance. J Clin Invest..
• James, J., Zemskova, M., Eccles, C. A., Varghese, M. V., Niihori, M., Barker, N. K., Luo, M., Mandarino, L. J., Langlais, P. R., Rafikova, O., & Rafikov, R. (2021). Single Mutation in the NFU1 Gene Metabolically Reprograms Pulmonary Artery Smooth Muscle Cells. Arteriosclerosis, Thrombosis, and Vascular Biology.
• Levine, A., Liktor-Busa, E., Lipinski, A. A., Couture, S., Balasubramanian, S., Aicher, S. A., Langlais, P. R., Vanderah, T. W., & Milnes, T. M. (2021). Sex differences in the expression of the endocannabinoid system within V1M cortex and PAG of Sprague Dawley rats. Biology of sex differences, 12(1), 60.
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  Several chronic pain disorders, such as migraine and fibromyalgia, have an increased prevalence in the female population. The underlying mechanisms of this sex-biased prevalence have yet to be thoroughly documented, but could be related to endogenous differences in neuromodulators in pain networks, including the endocannabinoid system. The cellular endocannabinoid system comprises the endogenous lipid signals 2-AG (2-arachidonoylglycerol) and AEA (anandamide); the enzymes that synthesize and degrade them; and the cannabinoid receptors. The relative prevalence of different components of the endocannabinoid system in specific brain regions may alter responses to endogenous and exogenous ligands.
• Mandarino, L. J., De Filippis, E. A., Grandjean, D., Luo, M., Coletta, D. K., Langlais, P. R., Finlayson, J., & Zapata Bustos, R. (2021). Altered Transcription Factor Expression Responses to Exercise in Insulin Resistance. Frontiers in Physiology.
• Polverino, F., Wu, T. D., Rojas-Quintero, J., Wang, X., Mayo, J., Tomchaney, M., Tram, J., Packard, S., Zhang, D., Cleveland, K., Cordoba-Lanus, E., Owen, C. A., Fawzy, A., Kinney, G. L., Hersh, C. P., Hansel, N. N., Doubleday, K., Sauler, M., Tesfaigzi, Y., , Ledford, J., et al. (2020). Metformin: Experimental and Clinical Evidence for a Potential Role in Emphysema Treatment. American Journal of Respiratory and Critical Care Medicine.
• Uhlorn, J. A., Husband, N. A., Romero-Aleshire, M. J., Moffett, C., Lindsay, M. L., Langlais, P. R., & Brooks, H. L. (2021). CD4+ T Cell-Specific Proteomic Pathways Identified in Progression of Hypertension Across Postmenopausal Transition. Journal of the American Heart Association.
• Zapata Bustos, R., Finlayson, J., Langlais, P. R., Coletta, D. K., Luo, M., Grandjean, D., De Filippis, E. A., & Mandarino, L. J. (2021). Altered Transcription Factor Expression Responses to Exercise in Insulin Resistance. Frontiers in Physiology.
• Zhang, H., Lipinski, A. L., Smith, A. F., Moutal, A., Khanna, R., Langlais, P. R., Milnes, T. M., & Vanderah, T. W. (2020). The effects of chronic morphine on the endogenous cannabinoid system in the ventral tegmental area. Frontiers In Pharmacology.
• van der Pijl, R. J., van den Berg, M., van de Locht, M., Shen, S., Bogaards, S. J., Conijn, S., Langlais, P. R., Chen, J., Hooijman, P. E., Labeit, S., Heunks, L. M., Granzier, H. L., & Ottenheijm, C. A. (2021). MARP1 (Muscle ankyrin repeat protein 1) locks titin to the sarcomeric thin filament and is a newly discovered passive force regulator. J Gen Physiol.
• Burt, J. M., Taylor, S. Z., Jacobsen, N. L., Pontifex, T. K., & Langlais, P. R. (2020). Serine 319 phosphorylation is necessary and sufficient to induce a Cx37 conformation that leads to arrested cell cycling.. J Cell Sci., 133. doi:doi:10.1242/jcs.240721
• Duron, D. I., Lei, W., Barker, N. K., Stine, C., Mishra, S., Blagg, B. S., Langlais, P. R., & Streicher, J. M. (2020). Inhibition of spinal cord Hsp90 enhances morphine anti-nociception by activating an ERK/RSK pathway.. Science Signaling.
• Harris, S., Langlais, P. R., Granger, K., Napierski, N., Touma, K., Moran, H., & Strom, J. (2020). A novel “cut and paste” method for in situ replacement of cMyBP-C reveals a new role for cMyBP-C in the regulation of contractile oscillations. Circulation Research.
• James, J., Varghese, M. V., Vasilyev, M., Langlais, P. R., Tofovic, S. P., Rafikova, O., & Rafikov, R. (2020). Complex III Inhibition-Induced Pulmonary Hypertension Affects the Mitochondrial Proteomic Landscape. International Journal of Molecular Sciences.
• Moinpour, M., Barker, N. K., Guzman, L. E., Jewett, J. C., Langlais, P. R., & Schwartz, J. C. (2020). Discriminating changes in protein structure using tyrosine conjugation. Protein Science.
• Pendleton, A. L., Antolic, A. T., Kelly, A. C., Davis, M. A., Camacho, L. E., Doubleday, K., Anderson, M. J., Langlais, P. R., Lynch, R. M., & Limesand, S. W. (2020). Lower oxygen consumption and Complex I activity in mitochondria isolated from skeletal muscle of fetal sheep with intrauterine growth restriction. American Journal of Physioly - Endocrinology & Metabolism.
• Jacobsen, N. L., Pontifex, T. K., Langlais, P. R., & Burt, J. M. (2019). Phosphorylation-dependent intra-domain interaction of the Cx37 carboxyl-terminus controls cell survival. Cancers.
• Krantz, J., Parker, S. S., Barker, N. K., Deer, C. G., Mouneimne, G., & Langlais, P. R. (2019). Insulin Induces Microtubule Stabilization and Regulates the Microtubule Plus-end Tracking Protein Network in Adipocytes.. Molecular & cellular proteomics : MCP.
• Luo, M., Willis, W. T., Coletta, D. K., Langlais, P. R., Mengos, A., Ma, W., Finlayson, J., Wagner, G. R., Baier, L. J., Nair, A., & Mandarino, L. J. (2019). Deletion of the Mitochondrial Protein VWA8 Induces Oxidative Stress and an HNF4α Compensatory Response in Hepatocytes.. Biochemistry.
• Pandey, R., Zhou, M., Islam, S., Chen, B., Langlais, P. R., Srivastava, A., Cooke, L. S., Weterings, E., Von Hoff, D., & Mahadevan, D. (2019). Carcinoembryonic antigen cell adhesion molecule 6 (CEACAM6) in Pancreatic Ductal Adenocarcinoma (PDA): An integrative analysis of a novel therapeutic target. Scientific Reports.
• Rafikov, R., McBride, M. L., Zemskova, M., Kurdyukov, S., McClain, N., Niihori, M., Langlais, P. R., & Rafikova, O. (2019). INOSITOL MONOPHOSPHATASE 1 (IMPA1) AS A NOVEL INTERACTING PARTNER OF RAGE IN PULMONARY HYPERTENSION.. American Journal of Physiology - Lung Cellular and Molecular Physiology.
• Tran, L., Langlais, P. R., Hoffman, N., Roust, L., & Katsanos, C. (2019). Mitochondrial ATP synthase β-subunit production rate and ATP synthase specific activity are reduced in skeletal muscle of humans with obesity.. Experimental Physiology.
• Kras, K. A., Langlais, P. R., Hoffman, N., Roust, L. R., Benjamin, T. R., De Filippis, E. A., Dinu, V., & Katsanos, C. S. (2018). Obesity modifies the stoichiometry of mitochondrial proteins in a way that is distinct to the subcellular localization of the mitochondria in skeletal muscle.. Metabolism.
• Rafikova, O., Williams, E. E., McBride, M. L., Zemskova, M., Srivastava, A., Nair, V., Desai, A., Langlais, P. R., Zemskov, E., Simon, M., Mandarino, L. J., & Rafikov, R. (2018). Hemolysis-induced lung vascular leakage contributes to the development of pulmonary hypertension. American Journal of Respiratory Cell and Molecular Biology.
• Suntravat, M., Langlais, P. R., Sanchez, E. E., & Nielsen, V. G. (2018). CatroxMP-II: a heme-modulated fibrinogenolytic metalloproteinase isolated from Crotalus atrox venom.. Biometals.
• Willis, W. T., Miranda-Grandjean, D., Hudgens, J., Willis, E. A., Finlayson, J., De Filippis, E. A., Zapata Bustos, R., Langlais, P. R., Mielke, C., & Mandarino, L. J. (2018). Dominant and sensitive control of oxidative flux by the ATP-ADP carrier in human skeletal muscle mitochondria: Effect of lysine acetylation.. Archives of Biochemistry and Biophysics.
• Kruse, R., Krantz, J., Barker, N., Coletta, R., Rafikov, R., Luo, M., Hoejlund, K., Mandarino, L. J., & Langlais, P. R. (2017). Characterization of the CLASP2 Protein Interaction Network Identifies SOGA1 as a Microtubule-Associated Protein. Molecular & cellular proteomics : MCP, 16(10), 1718-1735. doi:10.1074/mcp.RA117.000011
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  CLASP2 is a microtubule-associated protein that undergoes insulin-stimulated phosphorylation and co-localization with reorganized actin and GLUT4 at the plasma membrane. To gain insight to the role of CLASP2 in this system, we developed and successfully executed a streamlined interactome approach and built a CLASP2 protein network in 3T3-L1 adipocytes. Using two different commercially available antibodies for CLASP2 and an antibody for epitope-tagged, overexpressed CLASP2, we performed multiple affinity purification coupled with mass spectrometry (AP-MS) experiments in combination with label-free quantitative proteomics and analyzed the data with the bioinformatics tool Significance Analysis of Interactome (SAINT). We discovered that CLASP2 co-immunoprecipitates (co-IPs) the novel protein SOGA1, the microtubule-associated protein kinase MARK2, and the microtubule/actin-regulating protein G2L1. The GTPase-activating proteins AGAP1 and AGAP3 were also enriched in the CLASP2 interactome, although subsequent AGAP3 and CLIP2 interactome analysis suggests a preference of AGAP3 for CLIP2. Follow-up MARK2 interactome analysis confirmed reciprocal co-IP of CLASP2 and also revealed MARK2 can co-IP SOGA1, glycogen synthase, and glycogenin. Investigating the SOGA1 interactome confirmed SOGA1 can reciprocal co-IP both CLASP2 and MARK2 as well as glycogen synthase and glycogenin. SOGA1 was confirmed to colocalize with CLASP2 and also with tubulin, which identifies SOGA1 as a new microtubule-associated protein. These results introduce the metabolic function of these proposed novel protein networks and their relationship with microtubules as new fields of cytoskeleton-associated protein biology.
• Xie, X., Sinha, S., Yi, Z., Langlais, P. R., Madan, M., Bowen, B. P., Willis, W., & Meyer, C. (2017). Role of adipocyte mitochondria in inflammation, lipemia and insulin sensitivity in humans: effects of pioglitazone treatment. International journal of obesity (2005).
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  To gain further insight into the role of adipocyte mitochondria in systemic lipid metabolism, inflammation and insulin sensitivity in humans and to provide a better understanding of the mechanisms of action of the peroxisome proliferator-activated receptor gamma agonist pioglitazone.
• Campbell, L. E., Langlais, P. R., Day, S. E., Coletta, R. L., Benjamin, T. R., De, F., Madura, J., Mandarino, L. J., Roust, L. R., & Coletta, D. K. (2016). Identification of Novel Changes in Human Skeletal Muscle Proteome After Roux-en-Y Gastric Bypass Surgery. DIABETES, 65(9), 2724-2731.
• Kras, K. A., Willis, W. T., Barker, N., Czyzyk, T., Langlais, P. R., & Katsanos, C. S. (2016). Subsarcolemmal mitochondria isolated with the proteolytic enzyme nagarse exhibit greater protein specific activities and functional coupling. Biochemistry and biophysics reports, 6, 101-107.
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  Skeletal muscle mitochondria are arranged as a reticulum. Insight into the functional characteristics of such structure is achieved by viewing the network as consisting of "subsarcolemmal" (SS) and "intermyofibrillar" (IMF) regions. During the decades, most, but not all, published studies have reported higher (sometimes over 2-fold) enzyme and enzyme-pathway protein-specific activities in IMF compared to SS mitochondria. We tested the hypothesis that non-mitochondrial protein contamination might account for much of the apparently lower specific activities of isolated SS mitochondria. Mouse gastrocnemii (n = 6) were suspended in isolation medium, minced, and homogenized according to procedures typically used to isolate SS mitochondria. However, the supernatant fraction, collected after the first slow-speed (800×g) centrifugation, was divided equally: one sample was exposed to nagarse (MITO+), while the other was not (MITO-). Nagarse treatment reduced total protein yield by 25%, while it increased protein-specific respiration rates (nmol O2 min(-1) mg(-1)), by 38% under "resting" (state 4) and by 84% under maximal (state 3) conditions. Nagarse therefore increased the respiratory control ratio (state 3/state 4) by 30%. In addition, the ADP/O ratio was increased by 9% and the activity of citrate synthase (U/mg) was 49% higher. Mass spectrometry analysis indicated that the MITO+ preparation contained less contamination from non-mitochondrial proteins. We conclude that nagarse treatment of SS mitochondria removes not only non-mitochondrial proteins but also the protein of damaged mitochondria, improves indices of functional integrity, and the resulting protein-specific activities.
• Xie, X., Yi, Z., Sinha, S., Madan, M., Bowen, B. P., Langlais, P., Ma, D., Mandarino, L., & Meyer, C. (2016). Proteomics analyses of subcutaneous adipocytes reveal novel abnormalities in human insulin resistance. Obesity (Silver Spring, Md.), 24(7), 1506-14.
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  To provide a more global view of adipocyte changes in human insulin resistance by proteomics analyses.
• Xu, Q., Hou, Y., Langlais, P., Erickson, P., Zhu, J., Shi, C., Luo, M., Zhu, Y., Xu, Y. e., Mandarino, L. J., Stewart, K., & Chang, X. (2016). Expression of the cereblon binding protein argonaute 2 plays an important role for multiple myeloma cell growth and survival. BMC CANCER, 16.
• McLean, C. S., Mielke, C., Cordova, J. M., Langlais, P. R., Bowen, B., Miranda, D., Coletta, D. K., & Mandarino, L. J. (2015). Gene and MicroRNA Expression Responses to Exercise; Relationship with Insulin Sensitivity. PLOS ONE, 10(5).
• Aqel, B., Langlais, P., Vargas, H. E., Carey, E. J., Leonard, M., & Mandarino, L. J. (2014). Prospective Assessment of the Abundance of Drug Metabolizing Enzymes in the P450 Pathway in Patients with Non Alcoholic Fatty Liver Disease (NAFLD). HEPATOLOGY, 60, 759A-759A.
• Mielke, C., Lefort, N., McLean, C. G., Cordova, J. M., Langlais, P. R., Bordner, A. J., Te, J. A., Ozkan, S. B., Willis, W. T., & Mandarino, L. J. (2014). Adenine Nucleotide Translocase Is Acetylated in Vivo in Human Muscle: Modeling Predicts a Decreased ADP Affinity and Altered Control of Oxidative Phosphorylation. BIOCHEMISTRY, 53(23), 3817-3829.
• Xie, X., Langlais, P., Zhang, X., Heckmann, B. L., Saarinen, A. M., Mandarino, L. J., & Liu, J. (2014). Identification of a novel phosphorylation site in adipose triglyceride lipase as a regulator of lipid droplet localization. AMERICAN JOURNAL OF PHYSIOLOGY-ENDOCRINOLOGY AND METABOLISM, 306(12), E1449-E1459.
• Zhu, Y. X., Braggio, E., Shi, C., Kortuem, K. M., Bruins, L. A., Schmidt, J. E., Chang, X., Langlais, P., Luo, M., Jedlowski, P., LaPlant, B., Laumann, K., Fonseca, R., Bergsagel, P. L., Mikhael, J., Lacy, M., Champion, M. D., & Stewart, A. K. (2014). Identification of cereblon-binding proteins and relationship with response and survival after IMiDs in multiple myeloma. BLOOD, 124(4), 536-545.
• Chao, A., Zhang, X., Ma, D., Langlais, P., Luo, M., Mandarino, L. J., Zingsheim, M., Pham, K., Dillon, J., & Yi, Z. (2012). Site-specific phosphorylation of protein phosphatase 1 regulatory subunit 12A stimulated or suppressed by insulin. JOURNAL OF PROTEOMICS, 75(11), 3342-3350.
• Geetha, T., Langlais, P., Caruso, M., & Yi, Z. (2012). Protein phosphatase 1 regulatory subunit 12A and catalytic subunit delta, new members in the phosphatidylinositide 3 kinase insulin-signaling pathway. JOURNAL OF ENDOCRINOLOGY, 214(3), 437-443.
• Langlais, P., Dillon, J. L., Mengos, A., Baluch, D. P., Ardebili, R., Miranda, D. N., Xie, X., Heckmann, B. L., Liu, J., & Mandarino, L. J. (2012). Identification of a Role for CLASP2 in Insulin Action. JOURNAL OF BIOLOGICAL CHEMISTRY, 287(46), 39245-39253.
• Liu, M., Zhou, L., Wei, L. i., Villarreal, R., Yang, X., Hu, D., Riojas, R. A., Holmes, B. M., Langlais, P. R., Lee, H., & Dong, L. Q. (2012). Phosphorylation of Adaptor Protein Containing Pleckstrin Homology Domain, Phosphotyrosine Binding Domain, and Leucine Zipper Motif 1 (APPL1) at Ser(430) Mediates Endoplasmic Reticulum (ER) Stress-induced Insulin Resistance in Hepatocytes. JOURNAL OF BIOLOGICAL CHEMISTRY, 287(31), 26087-26093.
• Pham, K., Langlais, P., Zhang, X., Chao, A., Zingsheim, M., & Yi, Z. (2012). Insulin-stimulated phosphorylation of protein phosphatase 1 regulatory subunit 12B revealed by HPLC-ESI-MS/MS. PROTEOME SCIENCE, 10.
• Everman, S., Yi, Z., Langlais, P., Mandarino, L. J., Luo, M., Roberts, C., & Katsanos, C. S. (2011). Reproducibility of an HPLC-ESI-MS/MS Method for the Measurement of Stable-Isotope Enrichment of in Vivo-Labeled Muscle ATP Synthase Beta Subunit. PLOS ONE, 6(10).
• Geetha, T., Langlais, P., Luo, M., Mapes, R., Lefort, N., Chen, S., Mandarino, L. J., & Yi, Z. (2011). Label-Free Proteomic Identification of Endogenous, Insulin-Stimulated Interaction Partners of Insulin Receptor Substrate-1. JOURNAL OF THE AMERICAN SOCIETY FOR MASS SPECTROMETRY, 22(3), 457-466.
• Langlais, P., Yi, Z., & Mandarino, L. J. (2011). The Identification of Raptor as a Substrate for p44/42 MAPK. ENDOCRINOLOGY, 152(4), 1264-1273.
• Langlais, P., Mandarino, L. J., & Yi, Z. (2010). Label-free Relative Quantification of Co-eluting Isobaric Phosphopeptides of Insulin Receptor Substrate-1 by HPLC-ESI-MS/MS. JOURNAL OF THE AMERICAN SOCIETY FOR MASS SPECTROMETRY, 21(9), 1490-1499.
• Hojlund, K., Bowen, B. P., Hwang, H., Flynn, C. R., Madireddy, L., Geetha, T., Langlais, P., Meyer, C., Mandarino, L. J., & Yi, Z. (2009). In vivo Phosphoproteome of Human Skeletal Muscle Revealed by Phosphopeptide Enrichment and HPLC-ESI-MS/MS. JOURNAL OF PROTEOME RESEARCH, 8(11), 4954-4965.
• Hojlund, K., Yi, Z., Hwang, H., Bowen, B., Lefort, N., Flynn, C. R., Langlais, P., Weintraub, S. T., & Mandarino, L. J. (2008). Characterization of the human skeletal muscle proteome by one-dimensional gel electrophoresis and HPLC-ESI-MS/MS. MOLECULAR & CELLULAR PROTEOMICS, 7(2), 257-267.
• Hojlund, K., Yi, Z., Lefort, N., Langlais, P., Bowen, B., Levin, K., Beck-Nielsen, H., & Mandarino, L. J. (2008). Identification of multiple phosphorylation sites on ATP synthase beta subunit in human muscle. Increased phosphorylation at Thr213 and Tyr361 in obesity and type 2 diabetes. DIABETES, 57, A19-A19.
• Luo, M., Langlais, P., Yi, Z., Lefort, N., De Filippis, E. A., Hwang, H., Christ-Roberts, C. Y., & Mandarino, L. J. (2007). Phosphorylation of human insulin receptor substrate-1 at Serine 629 plays a positive role in insulin signaling. Endocrinology, 148(10), 4895-905.
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  The function of insulin receptor substrate-1 (IRS-1) is regulated by both tyrosine and serine/threonine phosphorylation. Phosphorylation of some serine/threonine residues in IRS-1 dampens insulin signaling, whereas phosphorylation of other serine/threonine residues enhances insulin signaling. Phosphorylation of human IRS-1 at Ser(629) was increased by insulin in Chinese hamster ovary cells expressing the insulin receptor (1.26 +/- 0.09-fold; P < 0.05) and L6 cells (1.35 +/- 0.29-fold; P < 0.05) expressing human IRS-1. Sequence analysis surrounding Ser(629) revealed conformity to the consensus phosphorylation sequence recognized by Akt. Phosphorylation of IRS-1 at Ser(629) in cells was decreased upon treatment with either an Akt inhibitor or by coexpression with kinase dead Akt, whereas Ser(629) phosphorylation was increased by coexpression with constitutively active Akt. In addition, Ser(629) of IRS-1 is directly phosphorylated by Akt in vitro. In cells, preventing phosphorylation of Ser(629) by a Ser(629)Ala mutation resulted in increased phosphorylation of Ser(636), a known negative regulator of IRS-1, without affecting phosphorylation of Tyr(632) or Ser(616). Cells expressing the Ser(629)Ala mutation, along with increased Ser(636) phosphorylation, had decreased insulin-stimulated association of the p85 regulatory subunit of phosphatidylinositol 3'-kinase with IRS-1 and decreased phosphorylation of Akt at Ser(473). Finally, in vitro phosphorylation of a Ser(629)-containing IRS-1 fragment with Akt reduces the subsequent ability of ERK to phosphorylate Ser(636/639). These results suggest that a feed-forward mechanism may exist whereby insulin activation of Akt leads to phosphorylation of IRS-1 at Ser(629), resulting in decreased phosphorylation of IRS-1 at Ser(636) and enhanced downstream signaling. Understanding the complex phosphorylation patterns of IRS-1 is crucial to elucidating the factors contributing to insulin resistance and, ultimately, the pathogenesis of type 2 diabetes.
• Luo, M., Langlais, P., Yi, Z., Lefort, N., De, F., Hwang, H., Christ-Roberts, C. Y., & Mandarino, L. J. (2007). Phosphorylation of human insulin receptor substrate-1 at serine 629 plays a positive role in insulin signaling. ENDOCRINOLOGY, 148(10), 4895-4905.
• Mapes, R., Yi, Z., Langlais, P., Riojas, R., Liu, F., Dong, L. Q., & Mandarino, L. J. (2007). APPL1, an adiponectin receptor binding partner, is phosphorylated at ser(401) in human skeletal muscle in vivo. DIABETES, 56, A508-A508.
• Yi, Z., Flynn, C. R., Langlais, P., Mandarino, L. J., & Hojlund, K. (2007). Comprehensive characterization of the human skeletal muscle proteome by LC-MS/MS. DIABETES, 56, A397-A397.
• Yi, Z., Langlais, P., De Filippis, E. A., Luo, M., Flynn, C. R., Schroeder, S., Weintraub, S. T., Mapes, R., & Mandarino, L. J. (2007). Global assessment of regulation of phosphorylation of insulin receptor substrate-1 by insulin in vivo in human muscle. Diabetes, 56(6), 1508-16.
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  Research has focused on insulin receptor substrate (IRS)-1 as a locus for insulin resistance. Tyrosine phosphorylation of IRS-1 initiates insulin signaling, whereas serine/threonine phosphorylation alters the ability of IRS-1 to transduce the insulin signal. Of 1,242 amino acids in IRS-1, 242 are serine/threonine. Serine/threonine phosphorylation of IRS-1 is affected by many factors, including insulin. The purpose of this study was to perform global assessment of phosphorylation of serine/threonine residues in IRS-1 in vivo in humans.
• Mao, X., Kikani, C. K., Riojas, R. A., Langlais, P., Wang, L., Ramos, F. J., Fang, Q., Christ-Roberts, C. Y., Hong, J. Y., Kim, R., Liu, F., & Dong, L. Q. (2006). APPL1 binds to adiponectin receptors and mediates adiponectin signalling and function. Nature cell biology, 8(5), 516-23.
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  Adiponectin, also known as Acrp30, is an adipose tissue-derived hormone with anti-atherogenic, anti-diabetic and insulin sensitizing properties. Two seven-transmembrane domain-containing proteins, AdipoR1 and AdipoR2, have recently been identified as adiponectin receptors, yet signalling events downstream of these receptors remain poorly defined. By using the cytoplasmic domain of AdipoR1 as bait, we screened a yeast two-hybrid cDNA library derived from human fetal brain. This screening led to the identification of a phosphotyrosine binding domain and a pleckstrin homology domain-containing adaptor protein, APPL1 (adaptor protein containing pleckstrin homology domain, phosphotyrosine binding (PTB) domain and leucine zipper motif). APPL1 interacts with adiponectin receptors in mammalian cells and the interaction is stimulated by adiponectin. Overexpression of APPL1 increases, and suppression of APPL1 level reduces, adiponectin signalling and adiponectin-mediated downstream events (such as lipid oxidation, glucose uptake and the membrane translocation of glucose transport 4 (GLUT4)). Adiponectin stimulates the interaction between APPL1 and Rab5 (a small GTPase) interaction, leading to increased GLUT4 membrane translocation. APPL1 also acts as a critical regulator of the crosstalk between adiponectin signalling and insulin signalling pathways. These results demonstrate a key function for APPL1 in adiponectin signalling and provide a molecular mechanism for the insulin sensitizing function of adiponectin.
• Ramos, F. J., Langlais, P. R., Hu, D., Dong, L. Q., & Liu, F. (2006). Grb10 mediates insulin-stimulated degradation of the insulin receptor: a mechanism of negative regulation. American journal of physiology. Endocrinology and metabolism, 290(6), E1262-6.
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  Growth factor receptor-bound protein 10 (Grb10) is an adapter protein that interacts with a number of tyrosine-phosphorylated growth factor receptors, including the insulin receptor (IR). To investigate the role of Grb10 in insulin signaling, we generated cell lines in which the expression levels of Grb10 are either overexpressed by stable transfection or suppressed by RNA interference. We found that suppressing endogenous Grb10 expression led to increased IR protein levels, whereas overexpression of Grb10 led to reduced IR protein levels. Altering Grb10 expression levels had no effect on the mRNA levels of IR, suggesting that the modulation occurs at the protein level. Reduced IR levels were also observed in cells with prolonged insulin treatment, and this reduction was inhibited in Grb10-deficient cells. The insulin-induced IR reduction was greatly reversed by MG-132, a proteasomal inhibitor, but not by chloroquine, a lysosomal inhibitor. IR underwent insulin-stimulated ubiquitination in cells, and this ubiquitination was inhibited in the Grb10-suppressed cell line. Together, our results suggest that, in addition to inhibiting IR kinase activity by directly binding to the IR, Grb10 also negatively regulates insulin signaling by mediating insulin-stimulated degradation of the receptor.
• Riojas, R. A., Kikani, C. K., Wang, C., Mao, X., Zhou, L., Langlais, P. R., Hu, D., Roberts, J. L., Dong, L. Q., & Liu, F. (2006). Fine tuning PDK1 activity by phosphorylation at Ser163. The Journal of biological chemistry, 281(31), 21588-93.
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  3-Phosphoinositide-dependent protein kinase-1 (PDK1) mediates phosphorylation and activation of members of the AGC protein kinase family and plays an essential role in insulin signaling and action. However, whether and how PDK1 activity is regulated in cells remains largely uncharacterized. In the present study, we show that PDK1 undergoes insulin-stimulated and phosphatidylinositol 3-kinase-dependent phosphorylation at Ser244 in the activation loop and at a novel site: Ser163 in the hinge region between the two lobes of the kinase domain. Sequence alignment studies revealed that the residue corresponding to Ser163 of PDK1 in all other AGC kinases is glutamate, suggesting that a negative charge at this site may be important for PDK1 function. Replacing Ser163 with a negatively charged residue, glutamate, led to a 2-fold increase in PDK1 activity. Molecular modeling studies suggested that phosphorylated Ser163 may form additional hydrogen bonds with Tyr149 and Gln223. In support of this, mutation of Tyr149 to Ala is sufficient to reduce PDK1 activity. Taken together, our results suggest that PDK1 phosphorylation of Ser163 may provide a mechanism to fine-tune PDK1 activity and function in cells.
• Yi, Z., Luo, M., Reyna, S. M., Weintraub, S. T., Langlais, P., & Mandarino, L. J. (2006). Quantification of serine phosphorylation of IRS-1 by HPLC-ESI-MS/MS. DIABETES, 55, A305-A305.
• Langlais, P., Wang, C., Dong, L. Q., Carroll, C. A., Weintraub, S. T., & Liu, F. (2005). Phosphorylation of Grb10 by mitogen-activated protein kinase: identification of Ser150 and Ser476 of human Grb10zeta as major phosphorylation sites. Biochemistry, 44(24), 8890-7.
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  Grb10 is a Src-homology 2 (SH2) and Pleckstrin-homology (PH) domain-containing protein that binds to several autophosphorylated receptor tyrosine kinases including the insulin receptor (IR). Our previous studies showed that Grb10 underwent insulin-stimulated serine phosphorylation, yet the kinase(s) responsible for phosphorylation and the sites of the phosphorylation remain unknown. In this report, we show that Grb10 is a direct substrate of the p42/44 mitogen-activated protein kinase (MAPK). In addition, we found that inhibition of the MAPK signaling pathway reduced Grb10 phosphorylation in cells. Using site-directed mutagenesis, phosphopeptide mapping, and capillary HPLC-electrospray-tandem mass spectrometry analysis, we identified Ser(150), Ser(418), and Ser(476) of human Grb10zeta as MAPK-mediated in vitro phosphorylation sites. In vivo labeling and two-dimensional phosphopeptide mapping studies revealed that Ser(150) and Ser(476) of human Grb10zeta are phosphorylated in intact cells. Replacing Ser(150) and Ser(476) with alanines reduced the inhibitory effect of human Grb10zeta on insulin-stimulated IRS1 tyrosine phosphorylation. Taken together, our findings suggest that phosphorylation of the adaptor protein may provide a feedback inhibitory mechanism by which Grb10 regulates insulin signaling.
• Luo, M., Reyna, S., Wang, L., Yi, Z., Carroll, C., Dong, L. Q., Langlais, P., Weintraub, S. T., & Mandarino, L. J. (2005). Identification of insulin receptor substrate 1 serine/threonine phosphorylation sites using mass spectrometry analysis: regulatory role of serine 1223. Endocrinology, 146(10), 4410-6.
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  Insulin receptor substrate 1 (IRS-1), an intracellular substrate of the insulin receptor tyrosine kinase, also is heavily phosphorylated on serine and threonine residues, and several serine phosphorylation sites alter the function of IRS-1. Because of the large number of serine/threonine residues, position-by-position analysis of these potential phosphorylation sites by mutagenesis is difficult. To circumvent this, we have employed matrix-assisted laser desorption/ionization time-of-flight and HPLC-electrospray ionization tandem mass spectrometry techniques to scan for serine and threonine residues that are phosphorylated in full-length human IRS-1 ectopically expressed in cells using an adenoviral vector. This approach revealed 12 phosphorylation sites on serine or threonine residues, 10 of which were novel sites. Seven of these sites were in proline-directed motifs, whereas five were in arginine-directed sites. Sequence inspection suggested that phosphorylation of Ser1223 might alter the interaction of IRS-1 with the protein tyrosine phosphatase Src homology domain 2 (SH2)-containing phosphatase-2 (SHP-2). Mutation of Ser1223 to alanine to prevent phosphorylation resulted in increased association of SHP-2 with IRS-1, decreased insulin-stimulated tyrosine phosphorylation of IRS-1 in CHO/IR cells, and decreased insulin-stimulated association of the p85 regulatory subunit of phosphatidylinositol-3-kinase with IRS-1. This mutation had no effect on association of IRS-1 with the insulin receptor. Sequence analysis showed the Ser1223 region to be widely conserved evolutionarily. These data suggest that phosphorylation of Ser1223 dampens association of IRS-1 with SHP-2, thereby increasing net insulin-stimulated tyrosine phosphorylation.
• Langlais, P., Dong, L. Q., Ramos, F. J., Hu, D., Li, Y., Quon, M. J., & Liu, F. (2004). Negative regulation of insulin-stimulated mitogen-activated protein kinase signaling by Grb10. Molecular endocrinology (Baltimore, Md.), 18(2), 350-8.
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  Grb10 is a Pleckstrin homology and Src homology 2 (SH2) domain-containing protein that binds to the tyrosine-phosphorylated insulin receptor in response to insulin stimulation. Loss of Grb10 function in mice results in fetal and placental overgrowth; however, the molecular mechanism remains unknown. In the present study, we show that overexpression of Grb10 in Chinese hamster ovary cells expressing the insulin receptor or in 3T3-L1 adipocytes reduced insulin-stimulated phosphorylation of MAPK. Overexpression of Grb10 in rat primary adipocytes also inhibited insulin-stimulated phosphorylation of the MAPK downstream substrate Elk1. To determine the mechanism by which Grb10 inhibited insulin-stimulated MAPK signaling, we examined whether Grb10 affects the phosphorylation of MAPK upstream signaling components. We found that overexpression of Grb10 inhibited the insulin-stimulated phosphorylation of Shc, a positive regulator of the MAPK signaling pathway. The inhibitory effect was diminished when the SH2 domain of Grb10 was deleted. The negative role of Grb10 in insulin signaling was established by suppression of endogenous Grb10 by RNA interference in HeLa cells overexpressing the insulin receptor, which enhanced insulin-stimulated phosphorylation of MAPK, Shc, and Akt. Taken together, our findings suggest that Grb10 functions as a negative regulator in the insulin-stimulated MAPK signaling pathway. In addition, the inhibitory effect of Grb10 on the MAPK pathway is most likely due to a direct block of insulin-stimulated Shc tyrosine phosphorylation.
• Li, Y., Langlais, P., Gamper, N., Liu, F., & Shapiro, M. S. (2004). Dual phosphorylations underlie modulation of unitary KCNQ K(+) channels by Src tyrosine kinase. The Journal of biological chemistry, 279(44), 45399-407.
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  Src tyrosine kinase suppresses KCNQ (M-type) K(+) channels in a subunit-specific manner representing a mode of modulation distinct from that involving G protein-coupled receptors. We probed the molecular and biophysical mechanisms of this modulation using mutagenesis, biochemistry, and both whole-cell and single channel modes of patch clamp recording. Immunoprecipitation assays showed that Src associates with KCNQ2-5 subunits but phosphorylates only KCNQ3-5. Using KCNQ3 as a background, we found that mutation of a tyrosine in the amino terminus (Tyr-67) or one in the carboxyl terminus (Tyr-349) abolished Src-dependent modulation of heterologously expressed KCNQ2/3 heteromultimers. The tyrosine phosphorylation was much weaker for either the KCNQ3-Y67F or KCNQ3-Y349F mutants and wholly absent in the KCNQ3-Y67F/Y349F double mutant. Biotinylation assays showed that Src activity does not alter the membrane abundance of channels in the plasma membrane. In recordings from cell-attached patches containing a single KCNQ2/3 channel, we found that Src inhibits the open probability of the channels. Kinetic analysis was consistent with the channels having two discrete open times and three closed times. Src activity reduced the durations of the longest open time and lengthened the longest closed time of the channels. The implications for the mechanisms of channel regulation by the dual phosphorylations on both channel termini are discussed.
• McClung, J. P., Roneker, C. A., Mu, W., Lisk, D. J., Langlais, P., Liu, F., & Lei, X. G. (2004). Development of insulin resistance and obesity in mice overexpressing cellular glutathione peroxidase. Proceedings of the National Academy of Sciences of the United States of America, 101(24), 8852-7.
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  Insulin resistance, a hallmark of type 2 diabetes, is associated with oxidative stress. However, the role of reactive oxygen species or specific antioxidant enzymes in its development has not been tested under physiological conditions. The objective of our study was to investigate the impact of overexpression of glutathione peroxidase 1 (GPX1), an intracellular selenoprotein that reduces hydrogen peroxide (H(2)O(2)) in vivo, on glucose metabolism and insulin function. The GPX1-overexpressing (OE) and WT male mice (n = 80) were fed a selenium-adequate diet (0.4 mg/kg) from 8 to 24 weeks of age. Compared with the WT, the OE mice developed (P < 0.05) hyperglycemia (117 vs. 149 mg/dl), hyperinsulinemia (419 vs. 1,350 pg/ml), and elevated plasma leptin (5 vs. 16 ng/ml) at 24 weeks of age. Meanwhile, these mice were heavier (37 vs. 27 g, P < 0.001) and fatter (37% vs. 17% fat, P < 0.01) than the WT mice. At 30-60 min after an insulin challenge, the OE mice had 25% less (P < 0.05) of a decrease in blood glucose than the WT mice. Their insulin resistance was associated with a 30-70% reduction (P < 0.05) in the insulin-stimulated phosphorylations of insulin receptor (beta-subunit) in liver and Akt (Ser(473) and Thr(308)) in liver and soleus muscle. Here we report the development of insulin resistance in mammals with elevated expression of an antioxidant enzyme and suggest that increased GPX1 activity may interfere with insulin function by overquenching intracellular reactive oxygen species required for insulin sensitizing.
• Wick, K. R., Werner, E. D., Langlais, P., Ramos, F. J., Dong, L. Q., Shoelson, S. E., & Liu, F. (2003). Grb10 inhibits insulin-stimulated insulin receptor substrate (IRS)-phosphatidylinositol 3-kinase/Akt signaling pathway by disrupting the association of IRS-1/IRS-2 with the insulin receptor. The Journal of biological chemistry, 278(10), 8460-7.
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  Grb10 has been proposed to inhibit or activate insulin signaling, depending on cellular context. We have investigated the mechanism by which full-length hGrb10gamma inhibits signaling through the insulin receptor substrate (IRS) proteins. Overexpression of hGrb10gamma in CHO/IR cells and in differentiated adipocytes significantly reduced insulin-stimulated tyrosine phosphorylation of IRS-1 and IRS-2. Inhibition occurred rapidly and was sustained for 60 min during insulin stimulation. In agreement with inhibited signaling through the IRS/PI 3-kinase pathway, we found hGrb10gamma to both delay and reduce phosphorylation of Akt at Thr(308) and Ser(473) in response to insulin stimulation. Decreased phosphorylation of IRS-1/2 may arise from impaired catalytic activity of the receptor, since hGrb10gamma directly associates with the IR kinase regulatory loop. However, yeast tri-hybrid studies indicated that full-length Grb10 blocks association between IRS proteins and IR, and that this requires the SH2 domain of Grb10. In cells, hGrb10gamma inhibited insulin-stimulated IRS-1 tyrosine phosphorylation in a dose-dependent manner, but did not affect IR catalytic activity toward Tyr(972) in the juxtamembrane region and Tyr(1158/1162/1163) in the regulatory domain. We conclude that binding of hGrb10gamma to IR decreases signaling through the IRS/PI 3-kinase/AKT pathway by physically blocking IRS access to IR.
• Wick, M. J., Dong, L. Q., Hu, D., Langlais, P., & Liu, F. (2001). Insulin receptor-mediated p62dok tyrosine phosphorylation at residues 362 and 398 plays distinct roles for binding GTPase-activating protein and Nck and is essential for inhibiting insulin-stimulated activation of Ras and Akt. The Journal of biological chemistry, 276(46), 42843-50.
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  A GTPase-activating protein (GAP)-associated 60-kDa protein has been found to undergo rapid tyrosine phosphorylation in response to insulin stimulation. However, whether this protein is a direct in vivo substrate for the insulin receptor (IR) tyrosine kinase and whether the tyrosine phosphorylation plays a role in insulin signaling remain to be established. Here we show that the insulin-stimulated tyrosine phosphorylation of the GAP-associated protein, now identified as p62(dok), is inhibited by Grb10, an adaptor protein that binds directly to the kinase domain of the IR, both in vitro and in cells. Replacing Tyr(362) and Tyr(398) with phenylalanine greatly decreased the IR-catalyzed p62(dok) tyrosine phosphorylation in vitro, suggesting that these two residues are the major IR-mediated phosphorylation sites. However, mutations at Tyr(362) and Tyr(398) only partially blocked insulin-stimulated p62(dok) tyrosine phosphorylation in cells, indicating that p62(dok) is also a target for other cellular tyrosine kinase(s) in addition to the IR. Replacing Tyr(362) with phenylalanine abolished the interaction between p62(dok) and Nck. Mutations at Tyr(362/398) of p62(dok) disrupted the interaction between p62(dok) and GAP and decreased the inhibitory effect of p62(dok) on the insulin-stimulated activation of Ras and Akt, but not mitogen-activated protein kinase. Furthermore, the inhibitory effect of p62(dok) on Akt phosphorylation could be blocked by coexpression of a constitutively active Ras. Taken together, our findings indicate that p62(dok) is a direct substrate for the IR tyrosine kinase and that phosphorylation at Tyr(362) and Tyr(398) plays an essential role for p62(dok) to interact with its effectors and negatively regulate the insulin signaling pathway.
• Langlais, P., Dong, L. Q., Hu, D., & Liu, F. (2000). Identification of Grb10 as a direct substrate for members of the Src tyrosine kinase family. Oncogene, 19(25), 2895-903.
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  Treatment of cells with insulin and protein tyrosine phosphatase inhibitors such as vanadate and pervanadate resulted in the tyrosine phosphorylation of Grb10, a Src homology 2 (SH2) and pleckstrin homology domain-containing adaptor protein which binds to a number of receptor tyrosine kinases including the insulin receptor (IR). Although Grb10 binds directly to the kinase domain of the IR, our data show that Grb10 is not a direct substrate for the IR tyrosine kinase. Consistent with this finding, Grb10 tyrosine phosphorylation in cells was inhibited by herbimycin A, a relatively specific inhibitor for members of the Src tyrosine kinase family, and by the expression of dominant negative Src or Fyn. In addition, Grb10 tyrosine phosphorylation was stimulated by expression of constitutively active Src or Fyn in cells and by incubation with purified Src or Fyn in vitro. The insulin stimulated or Src/Fyn-mediated tyrosine phosphorylation in vivo was significantly reduced when Grb10 tyrosine 67 was changed to glycine. This mutant form of Grb10 bound with higher affinity to the IR in cells than that of the wild-type protein, suggesting that tyrosine phosphorylation of Grb10 may normally negatively regulate its binding to the IR. Our data show that Grb10 is a new substrate for members of the Src tyrosine kinase family and that the tyrosine phosphorylation of the protein may play a potential role in cell signaling processes mediated by these kinases. Oncogene (2000).
• Dong, L. Q., Zhang, R. B., Langlais, P., He, H., Clark, M., Zhu, L., & Liu, F. (1999). Primary structure, tissue distribution, and expression of mouse phosphoinositide-dependent protein kinase-1, a protein kinase that phosphorylates and activates protein kinase Czeta. The Journal of biological chemistry, 274(12), 8117-22.
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  Phosphoinositide-dependent protein kinase-1 (PDK1) is a recently identified serine/threonine kinase that phosphorylates and activates Akt and p70(S6K), two downstream kinases of phosphatidylinositol 3-kinase. To further study the potential role of PDK1, we have screened a mouse liver cDNA library and identified a cDNA encoding the enzyme. The predicted mouse PDK1 (mPDK1) protein contained 559 amino acids and a COOH-terminal pleckstrin homology domain. A 7-kilobase mPDK1 mRNA was broadly expressed in mouse tissues and in embryonic cells. In the testis, a high level expression of a tissue-specific 2-kilobase transcript was also detected. Anti-mPDK1 antibody recognized multiple proteins in mouse tissues with molecular masses ranging from 60 to 180 kDa. mPDK1 phosphorylated the conserved threonine residue (Thr402) in the activation loop of protein kinase C-zeta and activated the enzyme in vitro and in cells. Our findings suggest that there may be different isoforms of mPDK1 and that the protein is an upstream kinase that activates divergent pathways downstream of phosphatidylinositol 3-kinase.

Presentations

• Tabsh, K. K., Pendleton, A. L., Barker, N. K., Langlais, P. R., Limesand, S. W., & Hill, M. G. (2020, March/Spring). Confirmation of Inflammatory Biomarkers from Maternal Plasma in Women with Intrahepatic Cholestasis of Pregnancy. Society for Reproductive Investigation 67th Annual Scientific Meeting.
• Zapata Bustos, R., Langlais, P. R., Coletta, D. K., De Filippis, E. A., Grandjean, D. N., & Mandarino, L. J. (2020, June). Lower Response of Connective Tissue Growth Factor (CTGF) to Exercise Characterizes Insulin Resistant Muscle. American Diabetes Association 80th scientific sessions. Chicago, IL: American Diabetes Association.
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  Rocio Zapata-Bustos, Paul Langlais, Dawn Coletta, Elena A. De Filippis, Danielle Grandjean, Lawrence J. MandarinoLower Response of Connective Tissue Growth Factor (CTGF) to Exercise Characterizes Insulin Resistant Muscle
• Barker, N., Krantz, J., Parker, S., Mouneimne, G., & Langlais, P. R. (2018, June/Summer). Characterization of the G2L1 Interactome Leads to the Discovery that CLIP2, G2L1, and EB1 Undergo Insulin-Stimulated Phosphorylation. American Diabetes Association. Orlando, Florida.
• Kras, K., Langlais, P. R., Willis, W. T., Mandarino, L., De Filippis, E., Roust, L., & Katsanos, C. (2016, June). Differential enrichment of Sub-sarcolemmal mitochondria with individual proteins between lean and obese, insulin-resistant subjects. American Diabetes Association Annual Meeting.

Poster Presentations

• Gackle, M. E., Lu, E., Goodmanson, M. J., Zelms, K. R., Rojas, N. G., McGraw, M. B., Nash, J. C., Lasher, J. D., Amaro, A., Moberly, A. P., & Langlais, P. R. (2024, January). Assessing Candidate Proteins Identified by Quantitative Proteomics for a Role in Insulin-Stimulated Glucose Uptake. Undergraduate Biology Research Program.
• Grijalva, A., Benson, D., Fink, J., Zhang, X., Coletta, D. K., Langlais, P. R., Pandey, A., Mangiola, A., Koevary, J., Lancaster, J., & Goldman, S. (2023, November). Immune Modulation to Treat Ischemic Heart Failure. American Heart Association Annual Meeting.
• McGraw, M. B., Nash, J. C., Rojas, N. G., Amaro, A., Gackle, M. E., Goodmanson, M. J., Zelms, K. R., Moberly, A. P., & Langlais, P. R. (2023, April). New Components of the Insulin Signaling Pathway: Investigating the Role of Novel Proteins in GLUT4 Translocation. UA W.A. Franke Honors College Pinnacle Annual Poster Session.
• Nash, J. C., McGraw, M. B., Rojas, N. G., Amaro, A., Gackle, M. E., Goodmanson, M. J., Zelms, K. R., Moberly, A. P., & Langlais, P. R. (2023, January). Increased Adipocyte Differentiation Potentially Leading to Increased Expression of Proteins Involved in Glucose Transporter 4 Translocation. Undergraduate Biology Research Program.
• Odeneye, R. E., Lipinski, A. A., Coletta, D. K., & Langlais, P. R. (2023, August). Proteomic Profiling of Differentiated Myotubes Identifies Protein Candidates Potentially Involved in Insulin-Stimulated Glucose Uptake. Undergraduate Biology Research Program.
• Rojas, N. G., McGraw, M. B., Nash, J. C., Goodmanson, M. J., Amaro, A., Zelms, K. R., Gackle, M. E., Moberly, A. P., & Langlais, P. R. (2023, April). Proteomic Profiling of Mature White Adipocytes Identifies Protein Candidates Potentially Involved in Insulin-Stimulated Glucose Uptake. UA W.A. Franke Honors College Pinnacle Annual Poster Session.
• Hoyer-Kimura, C., Konhilas, J. P., Langlais, P. R., Pier, M., Salcedo, V., & Fricks, J. (2022, October). Estrogen Status in Menopause and Sex Differences  within the Gut Promote Functionally Distinct Changes. Arizona Physiological Society.
• Iannuzo, N., Langlais, P. R., Guerra, S., & Ledford, J. (2022, June). CC16 DEFICIENCY IMPACTS PULMONARY EPITHELIAL-DRIVEN RESPONSES DURING MYCOPLASMA PNEUMONIAE INFECTION.. Aspen Lung Conference.
• Miller, K., Liu, X., McSwain, M., Jaurgui, E., Langlais, P. R., & Craig, Z. R. (2022, June). Effects of a Phthalate Mixture on Hormone and Ovarian Antral Follicle Protein Abundance  . Gordon Research Conference - Environmental Endocrine Disruptors.
• Moberly, A. P., Krantz, J., Roman, M., Batty, S. R., Surber, E., Lee, N. Y., & Langlais, P. R. (2022, January). The Role of G2L1 in Microtubule and Actin-based Insulin-Stimulated Delivery of Glucose Transporter 4 to the Plasma Membrane. Undergraduate Biology Research Program.
• Victor, R. A., Lipinski, A. A., Langlais, P. R., & Ledford, J. (2022, June). Identifying novel phase separation proteins in E. coli and human cells using SEC-MS. RNA Society Meeting.
• Akram, A., Iannuzo, N., Langlais, P. R., & Ledford, J. (2021, July). CC16 Deficiency in Context of Early-life Infections and Epithelial-driven Responses. KEYS Conference.
• Batty, S. R., Roman, M., McCrary, D., Moberly, A. P., Lipinski, A. A., Barker, N. K., Krantz, J., & Langlais, P. R. (2021, January). Identification of a Role for G2L1 in Insulin Action. Undergraduate Biology Research Program.
• Moberly, A. P., McCrary, D., Lipinski, A. A., Roman, M., Batty, S. R., Kwak, E., Lee, N. Y., & Langlais, P. R. (2021, January). Isolation of Microtubules and Microtubule Associated Proteins through Subcellular Fractionation. Undergraduate Biology Research Program.
• Vizcarra, V. S., Barber, K. R., Franca-Solomon, G., Smith-Flint, A., Majuta, L., Langlais, P. R., Milnes, T. M., Vanderah, T. W., & Riegel, A. C. (2021, November). Targeted manipulation of PFC 5HT2A receptors and KV7 channels attenuates chronic neuropathic pain in rats.. Society for Neuroscience.
• Gabriel, K., Duron, D., Barker, N. K., Langlais, P. R., & Streicher, J. M. (2020, April/Spring). Does DUSP15 promote activation of ERK MAPK signaling after Hsp90 inhibition in the spinal cord to promote opioid anti-nociception?. Experimental Biology.
• Hurtado, K., Keresztes, A., Barker, N. K., Langlais, P. R., & Streicher, J. M. (2020, April/Spring). Does the Mu-Delta Opioid Receptor Heterodimer Repress Akt Kinase to Reduce Opioid Anti-Nociception?. Experimental Biology.
• James, J., Varghese, M. V., Niihori, M., Zemskova, M., Langlais, P. R., Rafikova, O., & Rafikov, R. (2020, May/Summer). The NFU1 G206C mutation metabolically reprograms pulmonary artery smooth muscle cells, promotes proliferation and apoptosis resistance. American Thoracic Society.
• Kraft, M., Francisco, D., Childress, B., Kimura, H., & Langlais, P. R. (2020, May/Summer). Significant Differences in Airway Epithelial Cell Phosphorylated Proteins are Present in Unstimulated Asthma Compared to Cells from Non-asthmatic Participants. American Thoracic Society.
• Pendleton, A. L., Davis, M. A., Kelly, A. C., Camacho, L. E., Anderson, M. J., Langlais, P. R., & Limesand, S. W. (2020, March/Spring). Decreased Mitochondrial Complex I Activity in Skeletal Muscle of Growth Restricted Ovine Fetuses with Placental Insufficiency. Society for Reproductive Investigation 67th Annual Scientific Meeting.
• Sieffert, M. M., Keresztes, A., Barker, N. K., Langlais, P. R., & Streicher, J. M. (2020, April/Spring). Investigation of the Signaling Mechanisms of the Mu-Delta Opioid Receptor Heterodimer. Experimental Biology.
• Zapata Bustos, R., Langlais, P. R., Coletta, D. K., De Filippis, E. A., Grandjean, D. N., & Mandarino, L. J. (2020, June/Summer). Lower Response of Connective Tissue Growth Factor (CTGF) to Exercise Characterizes Insulin Resistant Muscle. American Diabetes Association 80th scientific sessions. Chicago, IL: American Diabetes Association.
  More info

  Rocio Zapata-Bustos, Paul Langlais, Dawn Coletta, Elena A. De Filippis, Danielle Grandjean, Lawrence J. MandarinoLower Response of Connective Tissue Growth Factor (CTGF) to Exercise Characterizes Insulin Resistant Muscle
• Barker, N. K., Krantz, J. L., & Langlais, P. R. (2019, June/Summer). Characterization of Insulin-Regulated CLASP2 Phosphorylation. American Diabetes Association.
• Dy, A. B., Barker, N. K., Langlais, P. R., & Ledford, J. (2019, Sept/Fall). Identification and functional testing of a novel receptor for SP-A on eosinophils.. 2019 European Respiratory Society Congress.
• Dy, A. B., Langlais, P. R., & Ledford, J. (2019, August/Summer). Identification and functional testing of a novel receptor for SP-A on eosinophils.. Gordon Conference – Lung Development, Injury and Repair.
• James, J., Zemskova, M., Eccles, C. A., Varghese, M. V., Niihori, M., Barker, N. K., Langlais, P. R., Rafikova, O., & Rafikov, R. (2019, Novermber/Fall). A single mutation in NFU1 metabolically reprograms pulmonary artery smooth muscle cells and drives proliferation with apoptosis resistance. The Society for Redox Biology and Medicine's 26th Annual Conference.
• Keresztes, A., Olson, K., Hguyen, P., Barker, N. K., Liu, Z., Langlais, P. R., Hruby, V., & Streicher, J. M. (2019, July/Summer). The Mu-Delta Opioid Receptor Heterodimer Acts as a Negative Feedback Brake to Reduce Opioid Anti-Nociception by Repression of CaMKII and Src Signaling. International Narcotics Research Conference - 50TH ANNIVERSARY MEETING.
• Keresztes, A., Olson, K., Hguyen, P., Barker, N. K., Liu, Z., Langlais, P. R., Hruby, V., & Streicher, J. M. (2019, May/Summer). Mu-Delta Opioid Receptor Heterodimer Antagonists: Novel Ligands that Enhance Opioid Analgesia while Reducing Opioid Withdrawal. 2019 NIH Pain Consortium Symposium - Pain Across the Lifespan.
• Langlais, P. R., Parker, S. S., Krantz, J., Kwak, E., Barker, N. K., Deer, C. G., Lee, N. Y., & Mouneimne, G. (2019, June/Summer). What’s on the Tube? Microtubule-Associated Proteins, Microtubule Stabilization, and Insulin Action.. Federation of American Societies For Experimental Biology - The Regulation of Glucose Metabolism Conference.
• Pandey, R., Zhou, M., Islam, S., Chen, B., Langlais, P. R., Srivastava, A., Cooke, L. S., Weterings, E., Von Hoff, D., & Mahadevan, D. (2019, March/Spring). Targeting carcinoembryonic antigen cell adhesion molecule 6 (CEACAM6) reshapes the tumor-stroma in pancreatic ductal adenocarcinoma (PDA). American Association for Cancer Research Annual Meeting 2019.
• Tabsh, K. K., Barker, N. K., Langlais, P. R., & Hill, M. G. (2019, March/Spring). Proteomic profile of maternal plasma in women with intrahepatic cholestasis of pregnancy.. Society for Reproductive Investigation 66th Annual Scientific Meeting.
• Uhlorn, J. A., Husband, N. A., Romero-Aleshire, M. J., Moffett, C. K., Kelly, A. C., Langlais, P. R., Nikolich-Zugich, J., & Brooks, H. L. (2019, September/Fall). Transcriptomic and Proteomic Analysis of CD4+ T Cells to Identify Sex Differences in Angiotensin II Signaling Pathways.. American Heart Association – Hypertension 2019 Scientific Sessions.
• Haider, N., Langlais, P. R., Moly, P. K., Barker, N. K., & Zhou, W. (2018, May). Resistin induced Differential Global Protein Expression in Human Macrophage. Vascular Discovery 2018 Scientific Sessions.
• Husband, N. A., Moffett, C., Romero-Aleshire, M. J., Uhlorn, J. A., Uhrlaub, J., Nunez, F., Nikolich-Zugich, J., Langlais, P. R., & Brooks, H. L. (2018, April). Angiotensin II-Induced Hypertension in VCD-Treated Menopausal Female Mice Elicits Significant Changes to the Splenic CD4+ Cell Proteome. Experimental Biology.
• Lopez-Pier, M., Cannon, D., Langlais, P. R., & Konhilas, J. P. (2018, October/Fall). AMP activated Protein Kinase and Estrogen-depednent mechanisms underlying increased susceptibility to CVD during Menopause. University of Arizona Biomedical Engineering Student Poster Session.
• Rafikova, O., Mandarino, L. J., Zemskov, E., Langlais, P. R., Desai, A., Srivastava, A., & Rafikov, R. (2018, May). Free heme-mediated endothelial barrier dysfunction contributes to the development of pulmonary hypertension. American Thoracic Society.
  More info

  Our data indicate that PH patients have increased levels of free Hb in plasma that correlate with disease severity and progression. There is also a significant accumulation of free Hb and depletion of haptoglobin in the sugen/hypoxia rat model. In rats, perivascular edema was observed during first two weeks of PH concomitant with increased infiltration of inflammatory cells. In the cell culture model of HLMVECs, we found that not hemoglobin but free heme-induced endothelial permeability via activation of the p38/HSP27 signaling pathway. Indeed, the rat model also exhibited an increased activation of p38/HSP27 during the initial phase of PH. Surprisingly, despite the increased levels of hemolysis and heme-mediated signaling; there was no heme oxygenase-1 activation. This can be explained by observed destabilization of HIF1a during the first two weeks of PAH regardless of hypoxic conditions. We found that heme-mediated effects on endothelium, at least in part, depend on Heme Carrier Protein 1 (HCP-1) and pharmacological inhibition of HCP-1 by sulfasalazine reduced barrier disruptive potential of the heme. Sulfasalazine administration to sugen/hypoxia rats results in attenuation of PH by a reduction in vascular remodeling in the lungs as well as decreasing right heart hypertrophy.
• Rafikova, O., Rafikov, R., Langlais, P. R., & McBride, M. L. (2018, May). Receptor for Advanced Glycation Endproducts (RAGE) regulates metabolic reprogramming induced over-proliferation in pulmonary hypertension. American Thoracic Society.
  More info

  Our data indicate that PH patients have increased levels of free Hb in plasma that correlate with disease severity and progression. There is also a significant accumulation of free Hb and depletion of haptoglobin in the sugen/hypoxia rat model. In rats, perivascular edema was observed during first two weeks of PH concomitant with increased infiltration of inflammatory cells. In the cell culture model of HLMVECs, we found that not hemoglobin but free heme-induced endothelial permeability via activation of the p38/HSP27 signaling pathway. Indeed, the rat model also exhibited an increased activation of p38/HSP27 during the initial phase of PH. Surprisingly, despite the increased levels of hemolysis and heme-mediated signaling; there was no heme oxygenase-1 activation. This can be explained by observed destabilization of HIF1a during the first two weeks of PAH regardless of hypoxic conditions. We found that heme-mediated effects on endothelium, at least in part, depend on Heme Carrier Protein 1 (HCP-1) and pharmacological inhibition of HCP-1 by sulfasalazine reduced barrier disruptive potential of the heme. Sulfasalazine administration to sugen/hypoxia rats results in attenuation of PH by a reduction in vascular remodeling in the lungs as well as decreasing right heart hypertrophy.
• Satyal, M. K., Bowser, S., McMillan, R. P., Davy, B. M., Langlais, P. R., Helm, R. F., Davy, K. P., & Hulver, M. W. (2018, November/Fall). Short-Term High-Fat Diet Alters Human Skeletal Muscle ERK 1/2 Phosphorylation. The Obesity Society @ Obesity Week.
• Satyal, M. K., Krantz, J., Parker, S., & Langlais, P. R. (2018, September/Fall). Human skeletal muscle phosphoproteome response to high-fat feeding.. American Physiological Societey - Maryland.
• Vasilyev, M., Langlais, P. R., Rafikova, O., & Rafikov, R. (2018, Novermber/Fall). Inhibition of respiratory chain Complex III irreversibly changes mitochondria proteomic landscape. The Society for Redox Biology and Medicine's 25th Annual Conference.
• Barker, N., Krantz, J., & Langlais, P. R. (2017, November). Targeted Quantitative Phosphoproteomics Discovers a Link between the Cytoskeleton-Associated Proteins SOGA1, G2L1, AGAP3, EB1, CLIP2, & MARK2 and the Insulin Signaling Pathway. UA College of Medicine Junior Investigators Symposium.
• Katsanos, C. S., Kras, K. A., Langlais, P. R., Willis, W. T., De Filippis, E. A., & Roust, L. R. (2017, June). Effects of Obesity on Skeletal Muscle Biokgical Pathways Associated with Subsarcolemal Versus Intermyofibrillar Mitochondria Revealed by Proteomic Analysis. American Diabetes Association Annual Meeting.
• Langlais, P. R., & Mandarino, L. J. (2017, November). UA Department of Medicine Quantitative Proteomics Lab. UA College of Medicine Junior Investigators Symposium.
• Langlais, P. R., & Mandarino, L. J. (2017, October). UA Department of Medicine Quantitative Proteomics Lab. UA College of Medicine Innovation Symposium.
• McBride, M. L., Williams, E. R., Langlais, P. R., Mandarino, L. J., Rafikov, R., & Rafikova, O. (2017, June). Inositol Monophosphate 1 (IMPA1) And Rage Interaction: The Role Of Novel Proliferative Pathway In Pulmonary Hypertension. American Heart Association.
• Rafikov, R., Srivastava, A., Desai, A., Langlais, P. R., Zemskov, E., Mandarino, L. J., & Rafikova, O. (2017, June). Hemolysis-mediated vascular permeability in lungs contributes to the development of pulmonary hypertension. American Heart Association.
  More info

  Our data indicate that PH patients have increased levels of free Hb in plasma that correlate with disease severity and progression. There is also a significant accumulation of free Hb and depletion of haptoglobin in the sugen/hypoxia rat model. In rats, perivascular edema was observed during first two weeks of PH concomitant with increased infiltration of inflammatory cells. In the cell culture model of HLMVECs, we found that not hemoglobin but free heme-induced endothelial permeability via activation of the p38/HSP27 signaling pathway. Indeed, the rat model also exhibited an increased activation of p38/HSP27 during the initial phase of PH. Surprisingly, despite the increased levels of hemolysis and heme-mediated signaling; there was no heme oxygenase-1 activation. This can be explained by observed destabilization of HIF1a during the first two weeks of PAH regardless of hypoxic conditions. We found that heme-mediated effects on endothelium, at least in part, depend on Heme Carrier Protein 1 (HCP-1) and pharmacological inhibition of HCP-1 by sulfasalazine reduced barrier disruptive potential of the heme. Sulfasalazine administration to sugen/hypoxia rats results in attenuation of PH by a reduction in vascular remodeling in the lungs as well as decreasing right heart hypertrophy.
• Zapata Bustos, R., Finlayson, J., Langlais, P. R., De Filippis, E. A., & Mandarino, L. J. (2017, November). Global acetylome analysis of muscle reveals differentially lysine acetylated sites in obesity.. Junior Investigator Poster Forum. Tucson, Arizona.: College of Medicine, University of Arizona.
  More info

  Zapata-Bustos R, Finlayson J, Langlais P, De Filippis EA, Mandarino LJ. Global acetylome analysis of muscle reveals differentially lysine acetylated sites in obesity. Junior Investigator Poster Forum, November 17, 2017. Tucson, Arizona.
• Sangam, S., Gupta, A., Yilmaz, S., Yuan, J. X., Langlais, P. R., & Desai, A. A. (2018, May). Endothelial Targets of Sox17 in Pulmonary Arterial Hypertension: A Quantitative Proteomic Approach. American Thoracic Society.

Other Teaching Materials

• Langlais, P. R. (2018. 7 Lessons to Keep in Mind If You Are Considering a Career as an Academic in Discovery-based Research. University of Texas Health Science Center at San Antonio.