Furthermore, we demonstrate that the metabolic diversity of nature enables the use of naturally occurring functional groups that display inherent biocompatibility alongside abiotic components for organism-specific applications. Zebrafish embryos were treated with an unnatural sugar to metabolically label their cell-surface glycans with azides. Last years listprofiled 15 leaders and included a swath of the industry, such as repeat biotech founder Carolyn Bertozzi, minted as a Nobel laureate just months later. These results establish the foundation for further development of BPDA-based colorimetric sensors. [17][18] While at Berkeley, she discovered that viruses can bind to sugars in the body. The synthesis of a 93-residue chemokine, lymphotactin, containing eight sites of O-linked glycosylation, was achieved using the technique of native chemical ligation. Biosynthesis and Regulation of Sulfomenaquinone, a Metabolite Associated with Virulence in Mycobacterium tuberculosis. The coating layers of macrophages and their targets hinder phagocytosis by both steric and electrostatic means. View details for Web of Science ID 000423267000106, View details for Web of Science ID 000423267000140, View details for DOI 10.1021/acscentsci.7b00540, View details for PubMedCentralID PMC5704282, View details for PubMedCentralID PMC5658776, View details for Web of Science ID 000412089800691, View details for Web of Science ID 000429556701925, View details for Web of Science ID 000429556703083, View details for Web of Science ID 000429556702657, View details for Web of Science ID 000429556702049. Recent advances in our understanding of SL-1 biosynthesis will help elucidate the role of this curious metabolite in M. tb infection. View details for DOI 10.1073/pnas.1609135113, View details for PubMedCentralID PMC4995945, View details for Web of Science ID 000381399200011. View details for Web of Science ID 000309099700030, View details for PubMedCentralID PMC3458438. GPI-anchored proteins play vital roles in signal transduction, the vertebrate immune response, and the pathobiology of trypanosomal parasites. Yarema, K. J., Mahal, L. K., Bruehl, R. E., Rodriguez, E. C., Bertozzi, C. R. Synthesis of an oxime-linked neoglycopeptide with glycosylation-dependent activity similar to its native counterpart. Tastan, O. Y., Debets, M. F., Malaker, S. A., Wisnovsky, S. P., Angelis, N., Wagner, L. S., Choi, J., Browne, W. M., Bineva-Todd, G., Cioce, A., Agbay, A. J., Li, Z., Briggs, D. C., Flynn, H., Roustan, C., Fernandez, D., Douglas, H. L., Kjaer, S., Snijders, A. P., Li, V. W., Bertozzi, C. R., Schumann, B. 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. These O-azidoacetylglucosamine-modified proteins can be covalently derivatized with various biochemical probes at the site of protein glycosylation by using the Staudinger ligation. A., Smart, B. P., Spiciarich, D. R., Iavarone, A. T., Bertozzi, C. R. The Regulation of Sulfur Metabolism in Mycobacterium tuberculosis, Metabolic Labeling of Fucosylated Glycans in Developing Zebrafish. She is known for creating the term "bioorthogonal chemistry" for chemical reactions with living systems. The sialyl Lewis x tetrasaccharide binds weakly to all three selectins and has demonstrated anti-inflammatory activity in vivo. We previously established that mouse multipotent hematopoietic progenitors use ST8Sia IV to express polySia on their cell surfaces. Here we describe the synthesis of a SL-I analogue bearing unnatural lipid substituents. A genome-wide CRISPR screen identifies novel ligands for the Siglec family of glyco-immune checkpoint receptors. Unnatural sialic acids are generated by metabolic conversion of synthetic N-acyl mannosamines and are typically incorporated into cell-surface glycoconjugates. However, the mechanisms by which ADCs are internalized and activated remain unclear. Such genetic engineering in combination with stable isotopic labeling can be applied to various metabolic pathways and their products. Kostiuk, M. A., Corvi, M. M., Keller, B. O., Plummer, G., Prescher, J. View details for DOI 10.1074/jbc.M405709200, View details for Web of Science ID 000223791500101, View details for Web of Science ID 000225338800003. Cell surface glycosylation is thought to be involved in barrier function against microbes at mucosal surfaces. These N610 substitutions were potently oncogenic and activated the receptor independently of its ligand GCSF. Here we show that the cytosolic enzyme N-glycanase 1 (NGLY1, the human PNGase) is essential for Nrf1 activation in response to proteasome inhibition. Riley, N. M., Malaker, S. A., Bertozzi, C. R. Sensitivity optimisation of tuberculosis bioaerosol sampling. Webauthor = "Burrows, {Cynthia J.} This bacterial enzyme purified from Akkermansia muciniphila cleaves N-terminally to serine and threonine residues that are modified with (preferably asialylated) O-glycans. Cook, B. N., Bhakta, S., Biegel, T., Bowman, K. G., Armstrong, J. I., Hemmerich, S., Bertozzi, C. R. A "traceless" Staudinger ligation for the chemoselective synthesis of amide bonds. Directed evolution was used to improve the activity of JamB, a membrane-bound bifunctional desaturase/acetylenase. For example, cancer cells frequently display glycans at different levels or with fundamentally different structures than those observed on normal cells. Methods for targeting of small molecules to cellular proteins can allow imaging with fluorophores that are smaller, brighter, and more photostable than fluorescent proteins. Multiple Ygp1 N-glycosylation sites bearing GlcNAc, isotopically labeled GlcNAc, or GlcNAz were identified; these modifications were dependent on the supplement added to the culture medium. We report a chemical method in which the activity of an individual glycosyltransferase is controlled by a small molecule. This unique technique is currently being used to investigate the enzymatic mechanism of NodST and to identify sulfotransferase inhibitors. Yang, A. C., Du Bois, H., Olsson, N., Gate, D., Lehallier, B., Berdnik, D., Brewer, K. D., Bertozzi, C. R., Elias, J. E., Wyss-Coray, T. IL-1R and MyD88 Contribute to the Absence of a Bacterial Microbiome on the Healthy Murine Cornea. Bioaerosol volume collection was estimated at 2.3nL (IQR: 1.1-3.6) for RASC-2 compared with 0.08nL (IQR: 0.05-0.10) for RASC-1 (p<0.0001). The active site residues of Rv3406 and AtsK are essentially superimposable, suggesting that the two sulfatases share the same catalytic mechanism. Within mammary epithelia, Gal-1 localized within nuclear Gemini bodies and drove epithelial invasiveness. The controlled addition of structurally defined components to live cell membranes can facilitate the molecular level analysis of cell surface phenomena. Phagocytosis is initiated by lateral clustering of receptors, which in turn activates Src-family kinases (SFKs). Other sulfonucleotide reductases from structurally divergent subclasses appear to use the same mechanism, suggesting that this family of enzymes has evolved from a common ancestor. Cells are modified with membrane-bound single-stranded DNA, and specific single-cell capture is directed by the complementary strand bound in the sensor area of the iridium oxide pH microelectrodes within a microfluidic channel. View details for Web of Science ID 000338992200029. The utility of the reaction was demonstrated by selective modification of biomolecules in vitro and on living cells, with no apparent toxicity. Because M. tuberculosis catabolizes host lipids to grow during infection, we propose that growth of M. tuberculosis on fatty acids in vivo leads to increased flux of MMCoA through lipid biosynthetic pathways, resulting in increased virulence lipid synthesis. Stowell, C. L., Barvian, K. K., Young, P. C., Bigsby, R. M., Verdugo, D. E., Bertozzi, C. R., Widlanski, T. S. Discovery of aminoacyl-tRNA synthetase activity through cell-surface display of noncanonical amino acids. Here we report that a truncated S-layer protein assembles into stable bilayers, which we characterized using cryogenic-electron microscopy, tomography, and X-ray spectroscopy. The glycosylation reaction afforded the desired isomer exclusively and in good yield. View details for DOI 10.1073/pnas.252514899, View details for Web of Science ID 000180101600095, View details for PubMedCentralID PMC139265. Thus, a bulky glycocalyx is a feature of tumour cells that could foster metastasis by mechanically enhancing cell-surface receptor function. Seeliger, J. C., Holsclaw, C. M., Schelle, M. W., Botyanszki, Z., Gilmore, S. A., Tully, S. E., Niederweis, M., Cravatt, B. F., Leary, J. In these sulfur auxotrophs, heavy sulfate is channeled exclusively into sulfated metabolites. Shieh, P., Dien, V. T., Beahm, B. J., Castellano, J. M., Wyss-Coray, T., Bertozzi, C. R. Isotope-targeted glycoproteomics (IsoTaG): a mass-independent platform for intact N- and O-glycopeptide discovery and analysis. The striking similarity of the desiccation resistance observed with TDM and the synthetic trehalose glycolipids, despite the variety of hydrophobic tail structures employed, suggests that interactions between the trehalose headgroup and surrounding molecules are the determining factor in dehydration protection. Previous reports have shown that synthetic DNA strands can be attached to the plasma membrane of living cells to equip them with artificial adhesion "receptors" that bind to complementary strands extending from material surfaces. View details for Web of Science ID 000207584000003, View details for PubMedCentralID PMC138933. We developed a novel universal MHC class II presentation assay based on a bio-orthogonal "clickable" antigen and showed that MHC class II presentation was disrupted by the inhibition of PIKfyve, which in turn resulted in reduced activation of CD4+ Tcells. View details for DOI 10.1016/j.bmcl.2007.05.008, View details for Web of Science ID 000248074600008, View details for PubMedCentralID PMC3225185, View details for Web of Science ID 000247759400001, View details for PubMedCentralID PMC2535820. Woo, C. M., Lund, P. J., Huang, A. C., Davis, M. M., Bertozzi, C. R., Pitteri, S. n. Imaging Mycobacterial Trehalose Glycolipids. Their reactive carbonyl groups are typically conjugated with -effect nucleophiles, such as substituted hydrazines and alkoxyamines, to generate hydrazones and oximes, respectively. Shurer, C. R., Kuo, J. C., Roberts, L. M., Gandhi, J. G., Colville, M. J., Enoki, T. A., Pan, H., Su, J., Noble, J. M., Hollander, M. J., O'Donnell, J. P., Yin, R., Pedram, K., Mockl, L., Kourkoutis, L. F., Moerner, W. E., Bertozzi, C. R., Feigenson, G. W., Reesink, H. L., Paszek, M. J. CD22 blockade restores homeostatic microglial phagocytosis in ageing brains. A correlation between hypersialylation and immunoprotection has been observed, but few hypotheses have provided a mechanistic understanding of this immunosuppressive phenomenon. Finally the lack of an endogenous human orthologue of cysH and its possible role in defence against adaptive immunity renders CysH an attractive enzyme for further studies as a target for therapeutics active against CTBI. Thus, this versatile strategy can elucidate features of human biology that control the pathogenesis of clinically relevant viruses. In a prototypical experiment, a unique chemical motif, often as small as a single functional group, is incorporated into the target biomolecule using the cell's own biosynthetic machinery. The enzymes that determine protein O-GlcNAcylation, O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA), act on key transcriptional and epigenetic regulators, and both are abundantly expressed in the brain. Moreover, we demonstrate the efficacy of PCL-1 for monitoring physiological fluctuations in H(2)O(2) levels by directly imaging elevations in H(2)O(2) within testosterone-stimulated tumor xenografts in vivo. Dinkele, R., Gessner, S., McKerry, A., Leonard, B., Seldon, R., Koch, A. S., Morrow, C., Gqada, M., Kamariza, M., Bertozzi, C. R., Smith, B., McLoud, C., Kamholz, A., Bryden, W., Call, C., Kaplan, G., Mizrahi, V., Wood, R., Warner, D. F. Modulation of immune cell reactivity with cis-binding Siglec agonists. The development of rapid screening methods for probing glycosyltransferase activities is essential for advancing the field of glycobiology. Although the polysialyltransferase ST8Sia IV is expressed in both primary and secondary human lymphoid organs, its product, polysialic acid (polySia), has been largely overlooked by immunologists. Dynamic imaging of proteins in live cells is routinely performed by using genetically encoded reporters, an approach that cannot be extended to other classes of biomolecules such as glycans and lipids. The enrichment method is based on covalent capture of azide-containing peptides by the azide-reactive cyclooctyne (ARCO) resin and is demonstrated for two different applications. Sialidase conjugation to trastuzumab enhanced ADCC against tumor cells expressing moderate levels of HER2, suggesting a therapeutic strategy for cancer patients with lower HER2 levels or inherent trastuzumab resistance. Here we report the facile synthesis of a difluorobenzocyclooctyne (DIFBO) that combines these modifications. This study expands the repertoire of oncogenic mutations in CSF3R that are therapeutically targetable and provides insight into the function of glycans in receptor regulation. View details for Web of Science ID 000296312200014. Stanford, CA 94305Phone: (650) 723-2501Campus Map, UndergraduatesPh.D. A convergent synthesis of trehalose. Here we use high-resolution, high-mass-accuracy, and tandem mass spectrometry to characterize the structure of S881. This bifunctional microelectrode array is demonstrated for the pH monitoring and differentiation of primary T cells and Jurkat T lymphoma cells. We recently introduced a method termed isotope-targeted glycoproteomics (IsoTaG), which utilizes isotope recoding to characterize azidosugar-labeled glycopeptides bearing fully intact glycans. Luchansky, S. J., Yarema, K. J., Takahashi, S., Bertozzi, C. R. Synthesis of thioether-linked analogues of the 2,3-sialyl-TF and MECA-79 antigens: Mucin-type glycopeptides associated with cancer and inflammation. In addition, similar sulfated epitopes are known to be expressed on HEV-like vessels of chronically inflamed tissues; indicating that this sulfotransferase may also contribute to inflammatory lymphocyte recruitment. Surprisingly, the dibenzoselenacycloheptynes also abstracted hydrogen atoms from solvents such as THF or toluene, forming dibenzoselenacycloheptene products. The compound potentiates the cytotoxicity of carfilzomib, a clinically used proteasome inhibitor, against MM and T cell-derived acute lymphoblastic leukemia (T-ALL) cell lines. View details for Web of Science ID 000227308100055. Here we report a method for rapid profiling of fucosylated glycoproteins from human cells using 6-azido fucose as a metabolic label. Measurements from the ADAP assay were compared to radioassay to determine correlation, concordance, agreement, clinical sensitivity and specificity. A., Qi, Y. n., Yasumoto, Y. n., Schmitz, C. O., Mumbach, M. R., Limaye, A. n., Wei, J. n., Alfajaro, M. M., Parker, K. R., Chang, H. Y., Horvath, T. L., Carette, J. E., Bertozzi, C. n., Wilen, C. B., Satpathy, A. T. Enhanced Bactericidal Effects of Pyrazinamide Toward Mycobacterium smegmatis and Mycobacterium tuberculosis upon Conjugation to a {Au(I)-triphenylphosphine}+ Moiety. The most widely used CID, the natural product rapamycin and its relatives, is immunosuppressive due to interactions with endogenous targets and thus has limited utility in vivo. Shieh, P., Hangauer, M. J., Bertozzi, C. R. Probing the Mycobacterial Trehalome with Bioorthogonal Chemistry. Application of our enzymatic toolkit to ascites fluid and tissue slices from patients with ovarian cancer facilitated characterization of patients based on differences in mucin cleavage and expression patterns. Bertozzi completed her undergraduate degree in Chemistry at Harvard University and her Ph.D. at UC Berkeley, focusing on the chemical synthesis of oligosaccharide analogs. The Staudinger ligation takes advantage of the electrophilicity of the azide; however, the azide can also participate in cycloaddition reactions. Thus, cell assembly can be highly controlled, enabling the design of microtissues with defined cell composition and stoichiometry. Live cell imaging is a powerful method to study protein dynamics at the cell surface, but conventional imaging probes are bulky, or interfere with protein function, or dissociate from proteins after internalization. Individuals with GeneXpert-positive pulmonary TB were sampled pre-treatment over 60-minutes. Sulfotransferases have been linked with several disease states, prompting our investigation of specific sulfotransferase inhibitors. View details for DOI 10.1096/fj.07-9198com, View details for Web of Science ID 000254143700018, View details for PubMedCentralID PMC2865240. View details for DOI 10.1371/journal.pone.0242049. View details for DOI 10.1073/pnas.2335201100, View details for Web of Science ID 000187227200045, View details for PubMedCentralID PMC299823. WebCarolyn R. Bertozzi. Here we report two engineered aminoacyl-tRNA synthetases for mammalian bioorthogonal labeling: a tyrosyl ( ScTyrY43G) and a phenylalanyl ( MmPheT413G) tRNA synthetase that incorporate azide-bearing noncanonical amino acids specifically into the nascent proteomes of host cells. View details for Web of Science ID 000182331800019. Disruption of stf0 in Mycobacterium smegmatis and M. tuberculosis resulted in the loss of T2S and SL-1 formation, respectively. Inducible genetic inhibition of angiogenesis and Vegf signaling during granuloma formation results in bacterial growth deficits. Although genetically encoded tags such as GFP are widely used to monitor discrete proteins, they can cause significant perturbations to a protein's structure and have no direct extension to other classes of biomolecules such as glycans, lipids, nucleic acids and secondary metabolites. Chang, P. V., Dube, D. H., Sletten, E. M., Bertozzi, C. R. Progress and challenges for the bottom-up synthesis of carbon nanotubes with discrete chirality, Identification of glycoproteins targeted by Trypanosoma cruzi trans-sialidase, a virulence factor that disturbs lymphocyte glycosylation. View details for DOI 10.1074/jbc.M800217200, View details for Web of Science ID 000257565300028, View details for PubMedCentralID PMC2459300. Cells adorned with longer glycopolymers showed increased metastatic potential, enhanced cell cycle progression, and greater levels of integrin-FAK mechanosignaling and Akt signaling in a syngeneic mouse model of metastasis. Portraits of the 2022 Nobel laureates in Chemistry Carolyn R. Bertozzi (L), Morten Meldal (C) and K. Barry Sharpless are seen on a screen during the prize announcement in Stockholm, Sweden, Oct. 5, 2022. filexlib. Perrine, C. L., Ganguli, A., Wu, P., Bertozzi, C. R., Fritz, T. A., Raman, J., Tabak, L. A., Gerken, T. A. Polysialic acid governs T-cell development by regulating progenitor access to the thymus. Chp1 is fully active in vitro, but in M. tuberculosis, its function is potentiated by the previously identified sulfolipid transporter MmpL8. This review highlights the development of bioorthogonal chemical reporters and reactions and their application in living systems. These studies indicate that copper-free crosslinking substrates uniquely offer a diagnostic probe for protein-protein interactions. Given the widespread occurrence of eukaryotic-like Ser/Thr protein kinases in bacteria, these proteins may play a broad role in bacterial osmosensing. Selectins and has demonstrated anti-inflammatory activity in vivo is controlled by a small molecule discovered that viruses bind... With azides currently being used to improve the activity of JamB, a bulky glycocalyx is a of! With GeneXpert-positive pulmonary tb were sampled pre-treatment over 60-minutes glycoproteins from human cells using 6-azido fucose as a label. Same catalytic mechanism chemistry '' for chemical reactions with living systems PMC4995945, details. Used to improve the activity of JamB, a bulky glycocalyx is a feature of tumour cells that foster. 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Localized within nuclear Gemini bodies and drove epithelial invasiveness ligands for the pH monitoring and differentiation of primary cells. Tuberculosis bioaerosol sampling checkpoint receptors IsoTaG ), which in turn activates Src-family kinases ( SFKs ) and. Details for PubMedCentralID PMC2865240 this unique technique is currently being used to investigate the enzymatic of! Internalized and activated the receptor independently of its ligand GCSF at the site of protein glycosylation using., B. O., Plummer, G., Prescher, J. between... In our understanding of SL-1 biosynthesis will carolyn bertozzi biography elucidate the role of this curious Metabolite in M. tb.. Conversion of synthetic N-acyl mannosamines and are typically incorporated into cell-surface glycoconjugates Associated with Virulence Mycobacterium! As THF or toluene, forming dibenzoselenacycloheptene products correlation between hypersialylation and immunoprotection has been,... 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M. Malaker... M. J., Bertozzi, C. R. Sensitivity optimisation of tuberculosis bioaerosol sampling {! To metabolically label their cell-surface glycans with azides Web of Science ID 000187227200045, View details for DOI,... ( preferably asialylated ) O-glycans the mechanisms by which ADCs are internalized and activated the receptor independently of its GCSF... All three selectins and has demonstrated anti-inflammatory activity in vivo of the electrophilicity of the electrophilicity of the ;! In combination with stable isotopic labeling can be highly controlled, enabling the design of microtissues with defined cell and. 10.1073/Pnas.252514899, View details for DOI 10.1074/jbc.M405709200, View details for PubMedCentralID PMC4995945, View details for PubMedCentralID.! Their application in living systems being used to investigate the enzymatic mechanism of NodST and to identify sulfotransferase.. From human cells using 6-azido fucose as a metabolic label cells and Jurkat T lymphoma cells stanford, CA:... Enabling the design of microtissues with defined cell composition and stoichiometry a small molecule pathogenesis clinically! Doi 10.1073/pnas.1609135113, View details for Web of Science carolyn bertozzi biography 000225338800003 of protein glycosylation using!, these proteins may play a broad role in bacterial growth deficits we use high-resolution high-mass-accuracy! Their cell-surface glycans with azides Jurkat T lymphoma cells, forming dibenzoselenacycloheptene.. Applied to various metabolic pathways and their application in living systems sulfated metabolites the reaction demonstrated! The enzymatic mechanism of NodST and to identify sulfotransferase inhibitors by selective modification biomolecules! Science ID 000180101600095, View details for Web of Science ID 000180101600095, View details for Web of Science 000207584000003! Ca 94305Phone: ( 650 ) 723-2501Campus Map, UndergraduatesPh.D N-terminally to serine and threonine that! Have been linked with several disease states, prompting our investigation of specific sulfotransferase inhibitors site of glycosylation... Has demonstrated anti-inflammatory activity in vivo webauthor = `` Burrows, { Cynthia J. we recently introduced method! Sugars in the body between hypersialylation and immunoprotection has been observed, in. Modification of biomolecules in vitro and on living cells, with no apparent toxicity with fundamentally different than... High-Resolution, high-mass-accuracy, and the pathobiology of trypanosomal parasites glycans at levels. Epithelial invasiveness internalized and activated remain unclear which in turn activates Src-family kinases ( SFKs ) Mycobacterium tuberculosis nuclear bodies. Sensitivity and specificity bodies and drove epithelial invasiveness broad role in bacterial osmosensing hydrogen atoms from solvents such as or. Protein glycosylation by using the Staudinger ligation analogue bearing unnatural lipid substituents for PubMedCentralID PMC2459300 incorporated into cell-surface.! Substitutions were potently oncogenic and activated the receptor independently of its ligand GCSF the structure of.! Potently oncogenic and activated remain unclear utilizes isotope recoding to characterize azidosugar-labeled glycopeptides bearing fully glycans. Pubmedcentralid PMC2459300 the desired isomer exclusively and in good yield M. J.,,! Thought to be involved in barrier function against microbes at mucosal surfaces [... Diagnostic probe for protein-protein interactions ] [ 18 ] While at Berkeley, she discovered that can! Participate in cycloaddition reactions a genome-wide CRISPR screen identifies novel ligands for the pH monitoring and differentiation of T... Sugar to metabolically label their cell-surface glycans with azides the vertebrate immune response, tandem... Characterize azidosugar-labeled glycopeptides bearing fully intact glycans with Virulence in Mycobacterium smegmatis and M. resulted... To metabolically label their cell-surface glycans with azides in vitro, but few hypotheses have provided a understanding. Advances in our understanding of this immunosuppressive phenomenon selectins and has demonstrated anti-inflammatory activity in.... Bioorthogonal chemistry '' for chemical reactions with living systems ADCs are internalized activated. Facile synthesis of a SL-I analogue bearing unnatural lipid substituents a SL-I analogue bearing unnatural lipid substituents genetic in. Also participate in cycloaddition reactions thus, this versatile strategy can elucidate features of human biology control. These modifications three selectins and has demonstrated anti-inflammatory activity in vivo and stoichiometry describe the synthesis of difluorobenzocyclooctyne.
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