John C Jewett

Interests

Teaching

organic chemistry, chemical biology

Research

Dengue virus, chemical biology, organic chemistry, virology

Courses

Scholarly Contributions

Journals/Publications

• 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.
• 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.
• Moinpour, M., Barker, N. K., Guzman, L. E., Jewett, J. C., Langlais, P. R., & Schwartz, J. C. (2020). Determining protein structure by tyrosine bioconjugation. ChemBioChem.
• Shadmehr, M., Davis, G. J., Mehari, B. T., Jensen, S. M., & Jewett, J. C. (2018). Coumarin Triazabutadienes for Fluorescent Labeling of Proteins. ChemBioChem, 19, 2550-2552. doi:10.1002/cbic.201800599
• He, J., Kimani, F. W., & Jewett, J. C. (2017). Rapid in Situ Generation of Benzene Diazonium Ions under Basic Aqueous Conditions from Bench-Stable Triazabutadienes. SYNLETT, 28(14), 1767-1770.
• Knyazeva, D. C., Kimani, F. W., Blanche, J., & Jewett, J. C. (2017). Hexyl triazabutadiene as a potent alkylating agent. TETRAHEDRON LETTERS, 58(28), 2700-2702.
• Martinez-Ariza, G., Mehari, B. T., Pinho, L., Foley, C., Day, K., Jewett, J. C., & Hulme, C. (2017). Synthesis of fluorescent heterocycles via a Knoevenagel/[4+1]-cycloaddition cascade using acetyl cyanide. ORGANIC & BIOMOLECULAR CHEMISTRY, 15(29), 6076-6079.
• Cornali, B. M., Kimani, F. W., & Jewett, J. C. (2016). Cu-Click Compatible Triazabutadienes To Expand the Scope of Aryl Diazonium Ion Chemistry. Organic letters, 18(19), 4948-4950.
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  Triazabutadienes can be used to readily generate reactive aryl diazonium ions under mild, physiologically relevant conditions. These conditions are compatible with a range of functionalities that do not tolerate traditional aryl diazonium ion generation. To increase the utility of this aryl diazonium ion releasing chemistry an alkyne-containing triazabutadiene was synthesized. The copper-catalyzed azide-alkyne cycloaddition ("Cu-click") reaction was utilized to modify the alkyne-containing triazabutadiene and shown to be compatible with the nitrogen-rich triazabutadiene. One of the triazole products was tethered to a fluorophore, thus enabling the direct fluorescent labeling of a model protein.
• Guzman, L. E., Kimani, F. W., & Jewett, J. C. (2016). Protecting Triazabutadienes To Afford Acid Resistance. Chembiochem : a European journal of chemical biology, 17(23), 2220-2222.
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  Recent work on triazabutadienes has shown that they have the ability to release aryl diazonium ions under exceptionally mild acidic conditions. There are instances that require that this release be prevented or minimized. Accordingly, a base-labile protection strategy for the triazabutadiene is presented. It affords enhanced synthetic and practical utility of the triazabutadiene. The effects of steric and electronic factors in the rate of removal are discussed, and the triazabutadiene protection is shown to be compatible with the traditional acid-labile protection strategy used in solid phase peptide synthesis.
• Jensen, S. M., Kimani, F. W., & Jewett, J. C. (2016). Light-Activated Triazabutadienes for the Modification of a Viral Surface. Chembiochem : a European journal of chemical biology, 17(23), 2216-2219.
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  Chemical crosslinking is a versatile tool for the examination of biochemical interactions, in particular host-pathogen interactions. We report the critical first step toward the goal of probing these interactions by the synthesis and use of a new heterobifunctional crosslinker containing a triazabutadiene scaffold. The triazabutadiene is stable to protein conjugation and liberates a reactive aryl diazonium species upon irradiation with 350 nm light. We highlight the use of this technology by modifying the surface of several proteins, including the dengue virus envelope protein.
• Jensen, S. M., Nguyen, C. T., & Jewett, J. C. (2016). A gradient-free method for the purification of infective dengue virus for protein-level investigations. Journal of virological methods, 235, 125-30.
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  Dengue virus (DENV) is a mosquito-transmitted flavivirus that infects approximately 100 million people annually. Multi-day protocols for purification of DENV reduce the infective titer due to viral sensitivity to both temperature and pH. Herein we describe a 5-h protocol for the purification of all DENV serotypes, utilizing traditional gradient-free ultracentrifugation followed by selective virion precipitation. This protocol allows for the separation of DENV from contaminating proteins - including intact C6/36 densovirus, for the production of infective virus at high concentration for protein-level analysis.
• He, J., Kimani, F. W., & Jewett, J. C. (2015). A Photobasic Functional Group. Journal of the American Chemical Society, 137(31), 9764-7.
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  Controlling chemical reactivity using light is a longstanding practice within organic chemistry, yet little has been done to modulate the basicity of compounds. Reported herein is a triazabutadiene that is rendered basic upon photoisomerization. The pH of an aqueous solution containing the water-soluble triazabutadiene can be adjusted with 350 nm light. Upon synthesizing a triazabutadiene that is soluble in aprotic organic solvents, we noted a similar light-induced change in basicity. As a proof of concept we took this photobase and used it to catalyze a condensation reaction.
• Kimani, F. W., & Jewett, J. C. (2015). Water-soluble triazabutadienes that release diazonium species upon protonation under physiologically relevant conditions. Angewandte Chemie (International ed. in English), 54(13), 4051-4.
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  Triazabutadienes are an understudied structural motif that have remarkable reactivity once rendered water-soluble. It is shown that these molecules readily release diazonium species in a pH-dependent manner in a series of buffer solutions with pH ranges similar to those found in cells. Upon further development, we expect that this process will be well suited to cargo-release strategies and organelle-specific bioconjugation reactions. These compounds offer one of the mildest ways of generating diazonium species in aqueous solutions.
• Ahad, A. M., Jensen, S. M., & Jewett, J. C. (2013). A traceless staudinger reagent to deliver diazirines. Organic Letters, 15(19), 5060-5063.
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  PMID: 24059816;PMCID: PMC3857746;Abstract: A triarylphosphine reagent that reacts with organic azides to install amide-linked diazirines is reported. This traceless Staudinger reagent reacts with complex organic azides to yield amide-linked diazirines, thus expanding the scope of the utility of both azide and diazirine chemistry. © 2013 American Chemical Society.
• Siegrist, M. S., Whiteside, S., Jewett, J. C., Aditham, A., Cava, F., & Bertozzi, C. R. (2013). D-amino acid chemical reporters reveal peptidoglycan dynamics of an intracellular pathogen. ACS Chemical Biology, 8(3), 500-505.
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  PMID: 23240806;PMCID: PMC3601600;Abstract: Peptidoglycan (PG) is an essential component of the bacterial cell wall. Although experiments with organisms in vitro have yielded a wealth of information on PG synthesis and maturation, it is unclear how these studies translate to bacteria replicating within host cells. We report a chemical approach for probing PG in vivo via metabolic labeling and bioorthogonal chemistry. A wide variety of bacterial species incorporated azide and alkyne-functionalized d-alanine into their cell walls, which we visualized by covalent reaction with click chemistry probes. The d-alanine analogues were specifically incorporated into nascent PG of the intracellular pathogen Listeria monocytogenes both in vitro and during macrophage infection. Metabolic incorporation of d-alanine derivatives and click chemistry detection constitute a facile, modular platform that facilitates unprecedented spatial and temporal resolution of PG dynamics in vivo. © 2012 American Chemical Society.
• Gordon, C. G., MacKey, J. L., Jewett, J. C., Sletten, E. M., Houk, K. N., & Bertozzi, C. R. (2012). Reactivity of biarylazacyclooctynones in copper-free click chemistry. Journal of the American Chemical Society, 134(22), 9199-9208.
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  PMID: 22553995;PMCID: PMC3368396;Abstract: The 1,3-dipolar cycloaddition of cyclooctynes with azides, also called "copper-free click chemistry", is a bioorthogonal reaction with widespread applications in biological discovery. The kinetics of this reaction are of paramount importance for studies of dynamic processes, particularly in living subjects. Here we performed a systematic analysis of the effects of strain and electronics on the reactivity of cyclooctynes with azides through both experimental measurements and computational studies using a density functional theory (DFT) distortion/interaction transition state model. In particular, we focused on biarylazacyclooctynone (BARAC) because it reacts with azides faster than any other reported cyclooctyne and its modular synthesis facilitated rapid access to analogues. We found that substituents on BARACs aryl rings can alter the calculated transition state interaction energy of the cycloaddition through electronic effects or the calculated distortion energy through steric effects. Experimental data confirmed that electronic perturbation of BARACs aryl rings has a modest effect on reaction rate, whereas steric hindrance in the transition state can significantly retard the reaction. Drawing on these results, we analyzed the relationship between alkyne bond angles, which we determined using X-ray crystallography, and reactivity, quantified by experimental second-order rate constants, for a range of cyclooctynes. Our results suggest a correlation between decreased alkyne bond angle and increased cyclooctyne reactivity. Finally, we obtained structural and computational data that revealed the relationship between the conformation of BARACs central lactam and compound reactivity. Collectively, these results indicate that the distortion/interaction model combined with bond angle analysis will enable predictions of cyclooctyne reactivity and the rational design of new reagents for copper-free click chemistry. © 2012 American Chemical Society.
• Swarts, B. M., Holsclaw, C. M., Jewett, J. C., Alber, M., Fox, D. M., Siegrist, M. S., Leary, J. A., Kalscheuer, R., & Bertozzi, C. R. (2012). Probing the mycobacterial trehalome with bioorthogonal chemistry. Journal of the American Chemical Society, 134(39), 16123-16126.
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  PMID: 22978752;PMCID: PMC3466019;Abstract: Mycobacteria, including the pathogen Mycobacterium tuberculosis, use the non-mammalian disaccharide trehalose as a precursor for essential cell-wall glycolipids and other metabolites. Here we describe a strategy for exploiting trehalose metabolic pathways to label glycolipids in mycobacteria with azide-modified trehalose (TreAz) analogues. Subsequent bioorthogonal ligation with alkyne-functionalized probes enabled detection and visualization of cell-surface glycolipids. Characterization of the metabolic fates of four TreAz analogues revealed unique labeling routes that can be harnessed for pathway-targeted investigation of the mycobacterial trehalome. © 2012 American Chemical Society.
• Dang, Y., Schneider-Poetsch, T., Eyler, D. E., Jewett, J. C., Bhat, S., Rawal, V. H., Green, R., & Liu, J. O. (2011). Inhibition of eukaryotic translation elongation by the antitumor natural product Mycalamide B. RNA, 17(8), 1578-1588.
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  PMID: 21693620;PMCID: PMC3153980;Abstract: Mycalamide B (MycB) is a marine sponge-derived natural product with potent antitumor activity. Although it has been shown to inhibit protein synthesis, the molecular mechanism of action by MycB remains incompletely understood. We verified the inhibition of translation elongation by in vitro HCV IRES dual luciferase assays, ribosome assembly, and in vivo [ 35S]methinione labeling experiments. Similar to cycloheximide (CHX), MycB inhibits translation elongation through blockade of eEF2-mediated translocation without affecting the eEF1A-mediated loading of tRNA onto the ribosome, AUG recognition, or dipeptide synthesis. Using chemical footprinting, we identified the MycB binding site proximal to the C3993 28S rRNA residue on the large ribosomal subunit. However, there are also subtle, but significant differences in the detailed mechanisms of action of MycB and CHX. First, MycB arrests the ribosome on the mRNA one codon ahead of CHX. Second, MycB specifically blocked tRNA binding to the E-site of the large ribosomal subunit. Moreover, they display different polysome profiles in vivo. Together, these observations shed new light on the mechanism of inhibition of translation elongation by MycB. Published by Cold Spring Harbor Laboratory Press. Copyright © 2011 RNA Society.
• Jewett, J. C., & Bertozzi, C. R. (2011). Synthesis of a fluorogenic cyclooctyne activated by Cu-free click chemistry. Organic Letters, 13(22), 5937-5939.
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  PMID: 22029411;PMCID: PMC3219546;Abstract: Cyclooctyne-based probes that become fluorescent upon reaction with azides are important targets for real-time imaging of azide-labeled biomolecules. The concise synthesis of a coumarin-conjugated cyclooctyne, coumBARAC, that undergoes a 10-fold enhancement in fluorescence quantum yield upon triazole formation with organic azides is reported. The design principles embodied in coumBARAC establish a platform for generating fluorogenic cyclooctynes suited for biological imaging. © 2011 American Chemical Society.
• Jewett, J. C., & Bertozzi, C. R. (2010). Cu-free click cycloaddition reactions in chemical biology. Chemical Society Reviews, 39(4), 1272-1279.
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  PMID: 20349533;PMCID: PMC2865253;Abstract: Bioorthogonal chemical reactions are paving the way for new innovations in biology. These reactions possess extreme selectivity and biocompatibility, such that their participating reagents can form covalent bonds within richly functionalized biological systems - in some cases, living organisms. This tutorial review will summarize the history of this emerging field, as well as recent progress in the development and application of bioorthogonal copper-free click cycloaddition reactions. © 2010 The Royal Society of Chemistry.
• Jewett, J. C., & Rawal, V. H. (2010). Temporary restraints to overcome steric obstacles: An efficient strategy for the synthesis of mycalamideB. Angewandte Chemie - International Edition, 49(46), 8682-8685.
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  PMID: 20931583;Abstract: Restrain and release: A one-pot MukaiyamaMichael/epoxidation sequence introduced three stereocenters, an intramolecular isocyanate trapping produced a rigid 10-membered cyclic carbamate, and the selective opening of the cyclic carbamate was used to reveal the fully constructed natural product. © 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
• Jewett, J. C., Sletten, E. M., & Bertozzi, C. R. (2010). Rapid Cu-free click chemistry with readily synthesized biarylazacyclooctynones. Journal of the American Chemical Society, 132(11), 3688-3690.
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  PMID: 20187640;PMCID: PMC2840677;Abstract: "Chemical equation presented" Bioorthogonal chemical reactions, those that do not interact or interfere with biology, have allowed for exploration of numerous biological processes that were previously difficult to study. The reaction of azides with strained alkynes, such as cyclooctynes, readily forms a triazole product without the need for a toxic catalyst. Here we describe a iarylazacyclooctynone (BARAC) that has exceptional reaction kinetics and whose synthesis is designed to be both modular and scalable. We employed BARAC for live cell fluorescence imaging of azide-labeled glycans. The high signal-to-background ratio obtained using nanomolar concentrations of BARAC obviated the need for washing steps. Thus, BARAC is a promising reagent for in vivo imaging. © 2010 American Chemical Society.
• Sletten, E. M., Nakamura, H., Jewett, J. C., & Bertozzi, C. R. (2010). Difluorobenzocyclooctyne: Synthesis, reactivity, and stabilization by β-cyclodextrin. Journal of the American Chemical Society, 132(33), 11799-11805.
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  PMID: 20666466;PMCID: PMC2923465;Abstract: Highly reactive cyclooctynes have been sought as substrates for Cu-free cycloaddition reactions with azides in biological systems. To elevate the reactivities of cyclooctynes, two strategies, LUMO lowering through propargylic fluorination and strain enhancement through fused aryl rings, have been explored. Here we report the facile synthesis of a difluorobenzocyclooctyne (DIFBO) that combines these modifications. DIFBO was so reactive that it spontaneously trimerized to form two asymmetric products that we characterized by X-ray crystallography. However, we were able to trap DIFBO by forming a stable inclusion complex with β-cyclodextrin in aqueous media. This complex could be stored as a lyophilized powder and then dissociated in organic solvents to produce free DIFBO for in situ kinetic and spectroscopic analysis. Using this procedure, we found that the rate constant for the cycloaddition reaction of DIFBO with an azide exceeds those for difluorinated cyclooctyne (DIFO) and dibenzocyclooctyne (DIBO). Cyclodextrin complexation is therefore a promising approach for stabilizing compounds that possess the high intrinsic reactivities desired for Cu-free click chemistry. © 2010 American Chemical Society.
• Gálvez, E., Romea, P., Urpí, F., Jewett, J. C., & Rawal, V. H. (2009). Preparation of (S)-4-Isopropyl-N-propanoyl-1,3-thiazolidine-2-thione. Organic Syntheses, 86, 70-80.
• Jewett, J. C., & Rawal, V. H. (2007). Total synthesis of pederin. Angewandte Chemie - International Edition, 46(34), 6502-6504.
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  PMID: 17645272;Abstract: (Chemical Equation Presented) Blisteringly fast: The potent cytotoxic blistering agent pederin has been synthesized (see scheme). The synthesis is diastereoselective and concise (just 12 steps for the longest linear sequence), and features a formal hetero-Diels-Alder reaction of a hindered diene, a Mukaiyama-Michael reaction to set two additional stereocenters, and a Curtius rearrangement to stereospecifically introduce the aminal functionality. © 2007 Wiley-VCH Verlag GmbH & Co. KGaA.
• Moncarz, J. R., Brunker, T. J., Jewett, J. C., Orchowski, M., Glueck, D. S., Sommer, R. D., Lam, K., Incarvito, C. D., Concolino, T. E., Ceccarelli, C., Zakharov, L. N., & Rheingold, A. L. (2003). Palladium-catalyzed asymmetric phosphination. Enantioselective synthesis of PAMP-BH₃, ligand effects on catalysis, and direct observation of the stereochemistry of transmetalation and reductive elimination. Organometallics, 22(16), 3205-3221.
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  Abstract: The complexes Pd(diphos)(o-An)(I) (o-An = o-MeOC6H4; diphos = dppe (3), (S,S)-Chiraphos (4), (R,R)-Me-Duphos (5), (R,S) -t-Bu-Josiphos (6), (R)-Tol-Binap (7)) were prepared. Complex 6 catalyzed the coupling of PH(Me)(Ph)(BH3) (2) with o-AnI in the presence of base to yield PAMP-BH3 (P(Me)(Ph)(o-An)(BH3) (1)) in low enantiomeric excess. The course of stoichiometric reactions of 3-7 with 2 and NaOSiMe3 depended on the diphosphine ligand. Complexes 6 and 7 gave PAMP-BH3 (1) and Pd(0) species; no intermediates were observed. With 3, the intermediate Pd(dppe)(o-An)(P(Me)(Ph)(BH3)) (10) was observed by 31P NMR, while 4 gave the isolable diastereomeric palladium complexes (Sp)-Pd((S,S)-Chiraphos)(o-An)(P(Me)(Ph)(BH3)) (11a) and (RP)-Pd((S,S)-Chiraphos)(o-An)(P(Me)(Ph)(BH3)) (11b), whose absolute configurations were determined by X-ray crystallography after separation. The analogous Pd((R,R)-Me-Duphos)(o-An)(P(Me)(Ph)(BH3)) diastereomers (12a,b) were also separated and isolated. Treatment of 4 with highly enantioenriched 2 (R or S) gave 11a or 11b in high diastereomeric excess with retention of configuration at phosphorus. P-C reductive elimination from either isomer of highly diastereoenriched 11 in the presence of excess diphenylacetylene yielded Pd((S,S)-Chiraphos)(PhC≡CPh) (14) and highly enantioenriched PAMP-BH3 (1), with retention of configuration.

Poster Presentations

• Jewett, J. C. (2014, June). Chemical Solutions to Viral Problems. Bioorganic Gordon Research Conference. Andover, NH.