carolyn bertozzi biography

The same mineralization technique, when applied to a hydrogel that is less prone to surface hydrolysis, led to distinctly different mineralization patterns, in terms of both the extent of mineralization and the crystallinity of the apatite grown on the hydrogel surface. Protein glycosylation is a heterogeneous post-translational modification (PTM) that plays an essential role in biological regulation. Both the activity and specificity of the inducible enzymes were indistinguishable from their WT counterparts. Individuals with GeneXpert-positive pulmonary TB were sampled pre-treatment over 60-minutes. Here, we report the synthesis and evaluation of a novel azacyclooctyne, 6,7-dimethoxyazacyclooct-4-yne (DIMAC). This type of enzyme catalyzes the initial step of mucin-type O-glycosylation, that is, the transfer of GalNAc in O-glycosidic linkage to serine and threonine residues in polypeptides. The structural identification of the fragments relied on the use of a variety of radiolabeled sugar precursors, further chemical and enzymatic hydrolysis, and high-pH anion-exchange chromatography analysis. These multivalent arrays are 4 orders of magnitude better than the monovalent carbohydrate. A., Laughlin, S. T., Agard, N. J., Chang, P. V., Miller, I. Her recent efforts include synthesis of chemical tools to study cell surface sugars called glycans and how they affect diseases such as cancer, inflammation, and viral infections like COVID-19. Together with determining their shedding and turnover rates, we also report that several membrane proteins, including TS and its substrates, both glycosylphosphatidylinositol-anchored proteins, are separately distributed on parasite surface and contained in different and highly stable membrane microdomains. The article will highlight recent approaches to thesynthesis of glycopeptide fragments bearing complex O-linkedglycans, as well as new strategies for the generation of full-lengthglycoproteins. Bacterial sulfate assimilation pathways provide for activation of inorganic sulfur for the biosynthesis of cysteine and methionine, through either adenosine 5'-phosphosulfate (APS) or 3'-phosphoadenosine 5'-phosphosulfate (PAPS) as intermediates. We previously described a chemical method to image glycans during zebrafish larval development; however, we were unable to detect glycans during the first 24 hours of embryogenesis, a very dynamic period in development. Saxon, E., Armstrong, J. I., Bertozzi, C. R. Cell surface engineering by a modified Staudinger reaction, Tyrosine sulfation: a modulator of extracellular protein-protein interactions, Discovery of carbohydrate sulfotransferase inhibitors from a kinase-directed library. Phagocytosis is the central process by which macrophage cells internalize and eliminate infectious microbes as well as apoptotic cells. A., Bertozzi, C. R. Metabolic selection of glycosylation defects in human cells, Polymerized liposome assemblies: Bifunctional macromolecular selectin inhibitors mimicking physiological selectin ligands. Lim, H. C., Sher, J. W., Rodriguez-Rivera, F. P., Fumeaux, C. n., Bertozzi, C. R., Bernhardt, T. G. The mucin-selective protease StcE enables molecular and functional analysis of human cancer-associated mucins. The purified enzyme catalyzed the sequential esterification of trehalose with two palmitoyl groups, generating a diacylated product similar to the 2,3-diacyltrehalose glycolipids of M. tuberculosis. The Staudinger ligation is based on the classic Staudinger reduction of azides with triarylphosphines first reported in 1919. [30] In 2015, Bertozzi moved to Stanford University to join the ChEM-H Institute. View details for DOI 10.1073/pnas.1608069113. She coined the term bioorthogonal chemistry to describe the use of click reactionsquick, simple chemical reactionsto study living cells. Debets, M. F., Tastan, O. Y., Wisnovsky, S. P., Malaker, S. A., Angelis, N. n., Moeckl, L. K., Choi, J. n., Flynn, H. n., Wagner, L. J., Bineva-Todd, G. n., Antonopoulos, A. n., Cioce, A. n., Browne, W. M., Li, Z. n., Briggs, D. C., Douglas, H. L., Hess, G. T., Agbay, A. J., Roustan, C. n., Kjaer, S. n., Haslam, S. M., Snijders, A. P., Bassik, M. C., Moerner, W. E., Li, V. S., Bertozzi, C. R., Schumann, B. n. Delaveris, C. n., Wilk, A. n., Riley, N. n., Stark, J. n., Yang, S. n., Rogers, A. n., Ranganath, T. n., Nadeau, K. n., Blish, C. n., Bertozzi, C. n. Systematic discovery and functional interrogation of SARS-CoV-2 viral RNA-host protein interactions during infection. The implications of the binary and ternary complexes observed by gas-phase noncovalent interactions in the mechanism of APS reduction are discussed. We developed a polymer coating for carbon nanotubes (CNTs) that mimics the mucin glycoprotein coating of mammalian cells. Loss of Stf3 expression, but not that of Cyp128, is correlated with elevated levels of menaquinone-9, an essential component in the electron-transport chain in M. tuberculosis. Moreover, approximately 1.3 microM total exogenous sialic acid was sufficient to obtain about 50% of the maximum production of sialic acid-containing glycoforms observed under in vitro growth conditions. Here, we report the development of antibody-sialidase conjugates that enhance tumor cell susceptibility to antibody-dependent cell-mediated cytotoxicity (ADCC) by selective desialylation of the tumor cell glycocalyx. The structure of sialic acid on living cells can be modulated by metabolism of unnatural biosynthetic precursors. Inactivation of the conserved steAB genes (cgp_1603-1604) was also found to confer EMB hypersensitivity and cell division defects. We have found that TDM monolayers, in stark contrast to phospholipid membranes, can be dehydrated and rehydrated without loss of integrity, as assessed by fluidity and protein binding. The data suggest that the ppGalNAcTs can be classified into at least four types, which working together, are able to produce densely glycosylated mucin glycoproteins. View details for DOI 10.1021/acs.analchem.0c02950. This provides orthogonal cleavage relative to canonical proteases (e.g., trypsin) for improved O-glycopeptide characterization with tandem mass spectrometry (MS/MS). Here we show that galectin-1 (Gal-1), an endogenous lectin that recognizes glycans bearing N-acetyllactosamine (LacNAc) epitopes, induces branching migration of mammary epithelia in vivo, ex vivo, and in 3D organotypic cultures. Each enzyme preferred a terminal GlcNAc residue, and was impeded by the addition of a beta1,4-linked Gal residue (i.e., terminal LacNAc). However, the copper catalysts are too cytotoxic for long-term exposure with live cells or organisms. Riley, N. M., Bertozzi, C. R., Pitteri, S. J. Little is known about how cells respond on a molecular level to tissue engineering scaffold materials. Ahn, G., Banik, S. M., Miller, C. L., Riley, N. M., Cochran, J. R., Bertozzi, C. R. Degradation from the outside in: targeting extracellular and membrane proteins for degradation through the endolysosomal pathway. View details for Web of Science ID 000257629200046, View details for PubMedCentralID PMC2664610. She has been awarded the Lemelson-MIT Prize, the Heinrich Wieland Prize, and a MacArthur Foundation Fellowship, among many others. Song, J., Chen, J., Klapperich, C. M., Eng, V., Bertozzi, C. R. A metabolic labeling approach toward proteomic analysis of mucin-type O-linked glycosylation. View details for Web of Science ID 000321093800027. View details for DOI 10.1128/AAC.01639-16, View details for PubMedCentralID PMC5328571. These findings enhance our understanding of mycobacterial cell envelope structure and dynamics and have implications for development of TB drug cocktails. Most identified proteins were involved in mechanisms of cellular metabolic functions, post-translational modifications, and some have been reported to be implicated in EOC etiology. Accordingly, treatment of cells with N-azidoacetylglucosamine results in the metabolic incorporation of the azido sugar into nuclear and cytoplasmic proteins. L-Selectin, a receptor bearing a C-type lectin domain, mediates the initial attachment of lymphocytes to high endothelial venules of lymph nodes. Sogi, K. M., Holsclaw, C. M., Fragiadakis, G. K., Nomura, D. K., Leary, J. Changes in O-linked protein glycosylation are known to correlate with disease states but are difficult to monitor in a physiological setting because of a lack of experimental tools. Using these probes, we studied the mobilities of labeled glycolipids by time-lapse microscopy and fluorescence recovery after photobleaching experiments and found that mycomembrane fluidity varies widely across species and correlates with mycolic acid structure. This approach is compatible with a wide range of cell types, offers excellent capture efficiency, and can potentially be used to create complex multicellular arrangements through the use of multiple capture sequences. The efficacy of antimicrobial drugs against Mycobacterium tuberculosis, an intracellular bacterial pathogen, is generally first established by testing compounds against bacteria in axenic culture. We find the S-layer-on-mica system possesses a kinetic trap associated with conformational differences between a long-lived transient state and the final stable state. Interestingly, two sulfotransferase genes are highly similar to mammalian sulfotransferases, increasing the list of mycobacterial eukaryotic-like protein families. Bertozzi is a member of the Royal Society and the academies of sciences of Germany and the United States. Among her many honours are the Lemelson-MIT Prize (2010), the Arthur C. Cope Award of the American Chemical Society (2017), and the Wolf Prize in Chemistry (2022). This article contains raw and processed data related to research published in "Role of the polypeptide N-acetylgalactosaminyltransferase 3 in ovarian cancer progression: possible implications in abnormal mucin O-glycosylation" [1]. Our results reveal that these transferases have unique peptide and glycopeptide preferences demonstrating their substrate diversity and their likely roles ranging from initiating transferases to filling-in transferases. Using a multicolor detection strategy, we performed a spatiotemporal analysis of glycan expression and trafficking and identified patterns that would be undetectable with conventional molecular imaging approaches. Agard, N. J., Baskin, J. M., Prescher, J. View details for DOI 10.1016/j.bmc.2005.04.085, View details for Web of Science ID 000231341900006. Williams, S. J., Senaratne, R. H., Mougous, J. D., Riley, L. W., Bertozzi, C. R. A 96-well dot-blot assay for carbohydrate sulfotransferases, Sulfotransferases and sulfatases in mycobacteria. The importance of sulfated molecules in cell-cell communication motivated us to develop a rapid two-step method for identifying these metabolites in microorganisms, particularly in pathogenic mycobacteria. Here we use high-resolution, high-mass-accuracy, and tandem mass spectrometry to characterize the structure of S881. Further, enzymatically simplified N-glycans are preserved on peptides. This left Bertozzi and the rest of the lab to complete their Ph.D. work with no direct supervision. Mycobacterial carbohydrate sulfotransferase Stf0 catalyzes the sulfuryl group transfer from 3'-phosphoadenosine-5'-phosphosulfate (PAPS) to trehalose. One such metabolite from M. tuberculosis lipid extracts, S881, has been shown to negatively regulate the virulence of M. tuberculosis in mouse infection studies, and its cell-surface localization suggests a role in modulating host-pathogen interactions. These receptors are promising targets for vaccine development and cancer immunotherapy. Putative Mtb bacilli were detected in collected fluid by fluorescence microscopy utilising DMN-Trehalose. Barnes, J., Kaushik, S., Bainer, R. O., Sa, J. K., Woods, E. C., Kai, F., Przybyla, L., Lee, M., Lee, H., Tung, J. C., Maller, O., Barrett, A. S., Lu, K. V., Lakins, J. N., Hansen, K. C., Obernier, K., Alvarez-Buylla, A., Bergers, G., Phillips, J. J., Nam, D., Bertozzi, C. R., Weaver, V. M. Glycosyltransferase bump-hole engineering to dissect mucin-type O-glycosylation in the living cell. We report a chemical approach for probing PG in vivo via metabolic labeling and bioorthogonal chemistry. 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. Marcaurelle, L. A., Pratt, M. R., Bertozzi, C. R. A new approach to mineralization of biocompatible hydrogel scaffolds: An efficient process toward 3-dimensional bonelike composites, Synthesis of a bisubstrate analogue targeting estrogen sulfotransferase. Our optimized two-step enzymatic/chemical labeling scheme was used to tag and image a variety of LAP fusion proteins in multiple mammalian cell lines with diverse fluorophores including fluorescein, rhodamine, Alexa Fluor 568, ATTO 647N, and ATTO 655. In vivo molecular imaging holds promise for understanding the underlying mechanisms of health, injury, aging, and disease, as it can detect distinct biochemical processes such as enzymatic activity, reactive small-molecule fluxes, or post-translational modifications. Furthermore, GalNAz-labeled B cells could be selectively targeted with a phosphine probe by Staudinger ligation within the living animal. Attachment via these "cell-adhesion barcodes" is rapid and specific, with close-packed arrays of cells forming within minutes. A., Krishnan, V. n., Pett, C. n., Yu, J. n., Woods, E. C., Kramer, J. R., Westerlind, U. n., Dorigo, O. n., Bertozzi, C. R. Quantitative Super-Resolution Microscopy of the Mammalian Glycocalyx. We also explore prospects for improving measurements to better regularize protein-level biology and efficiently associate PTMs to function and phenotype. Carrico, I. S., Carlson, B. L., Bertozzi, C. R. A cell nanoinjector based on carbon nanotubes. SiaNAz was found to comprise between 4% and 41% of total sialosides, depending on the system. We postulate that covalent modification and perhaps inhibition of various mitochondrial enzymes by palmitoyl-CoA could lead to the metabolic impairments found in obesity-related diseases. Selective chemical reactions that are orthogonal to the diverse functionality of biological systems have become important tools in the field of chemical biology. [7] In 2010, she was the first woman to receive the prestigious Lemelson-MIT Prize faculty award. Here we report the development of an HIV OF assay based on Antibody Detection by Agglutination-PCR (ADAP) technology. Although the protein products of these viruses have been studied in depth, how the RNA genomes operate inside human cells is poorly understood. These results suggest that glycopolymer microarrays can reveal discrete higher-order binding preferences beyond the recognition of individual glycan epitopes. We have demonstrated that the quadricyclane ligation is compatible with, and orthogonal to, strain-promoted azide-alkyne cycloaddition and oxime ligation chemistries by performing all three reactions in one pot on differentially functionalized protein substrates. Here, we demonstrate that, relative to wild-type controls, ST8Sia IV(-/-) mice have a 30% reduction in total thymocytes and a concomitant deficiency in the earliest thymocyte precursors. Thus, cell surface ketones are a potential vehicle for a metabolically controlled small-molecule drug delivery system. However, only some of these mutants were able to generate protection equivalent to that of BCG in mice. Higher order oligosaccharides were readily generated by alkylation of the corresponding 3-thioGalNAc with N-bromoacetamido sugars. In this context, polySia modulates cellular adhesion, migration, cytokine response, and contact-dependent differentiation. The accumulation of an SL-1 precursor, termed SL(1278), in mmpL8 mutant cells indicates that MmpL8 is necessary for an intermediate step in the SL-1 biosynthesis pathway. This strain-promoted azide-alkyne cycloaddition is often referred to as "Cu-free click chemistry". Mycobacteria are endowed with a highly impermeable mycomembrane that confers intrinsic resistance to many antibiotics. Cell surface glycosylation is thought to be involved in barrier function against microbes at mucosal surfaces. Fusion of these domains to the proteins FRB and FKBP enabled their induced assembly by the natural product rapamycin. View details for DOI 10.1529/biophysj.107.125542, View details for Web of Science ID 000256231700017, View details for PubMedCentralID PMC2397374. Bouchard, G., Garcia-Marques, F. J., Karacosta, L. G., Zhang, W., Bermudez, A., Riley, N. M., Varma, S., Mehl, L. C., Benson, J. Woods, E. C., Yee, N. A., Shen, J., Bertozzi, C. R. Glycocalyx Engineering with a Recycling Glycopolymer that Increases Cell Survival In Vivo. View details for DOI 10.1073/pnas.252514899, View details for Web of Science ID 000180101600095, View details for PubMedCentralID PMC139265. Mycobacterium tuberculosis (M. tuberculosis) is an intracellular pathogen possessing a complex mixture of cell wall lipids that are thought to modulate the activities of host macrophages. We engineered tumor cells to display glycocalyces of various thicknesses by coating them with synthetic mucin-mimetic glycopolymers. A., Bertozzi, C. R. A chemically synthesized version of the insect antibacterial glycopeptide, diptericin, disrupts bacterial membrane integrity. A., Qi, Y., Yasumoto, Y., Wei, J., Alfajaro, M. M., Shi, Q., Mumbach, M. R., Limaye, A., DeWeirdt, P. C., Schmitz, C. O., Parker, K. R., Woo, E., Chang, H. Y., Horvath, T. L., Carette, J. E., Bertozzi, C. R., Wilen, C. B., Satpathy, A. T. Author Correction: The clinical impact of glycobiology: targeting selectins, Siglecs and mammalian glycans. View details for DOI 10.1016/S0166-6851(03)00196-8, View details for Web of Science ID 000185793400002. The best reagent for this application was dependent upon the specific structure of the azide. We combined CRISPR-Cas9 knockout screens with RNAsequencing analysis to discover age-related genetic modifiers of microglial phagocytosis. CDG-Tre fluoresces upon activation by BlaC, the -lactamase uniquely expressed by Mtb, and the fluorescent product is subsequently incorporated within the bacterial cell wall via trehalose metabolic pathway. Using this knowledge, glycosyltransferase assembly lines have been redesigned for the production of specific glycan structures using protein engineering and chemical approaches. View details for DOI 10.1038/s41467-019-12684-7. Preliminary screening has identified compounds that inhibit estrogen sulfotransferase (EST), an enzyme relevant to breast cancer. Front-line tuberculosis (TB) drugs have been characterized extensively invitro and invivo with respect to gene expression and cell viability. Here, we report a Cu-free variant of click chemistry that can label these biomolecules rapidly and selectively in living systems, overcoming the intrinsic toxicity of the canonical Cu-catalyzed reaction. 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Targeted with a phosphine probe by Staudinger ligation is based on carbon nanotubes chemical.! Targeted with a phosphine probe by Staudinger ligation within the living animal the field of chemical biology assembly have... Cells with N-azidoacetylglucosamine results in the mechanism of APS reduction are discussed this strain-promoted azide-alkyne cycloaddition often... Into nuclear and cytoplasmic proteins mycobacterial eukaryotic-like protein families a potential vehicle for metabolically. She coined the term bioorthogonal chemistry to describe the use of click reactionsquick, simple chemical reactionsto study cells... Sianaz was found to confer EMB hypersensitivity and cell viability has been awarded the Lemelson-MIT Prize faculty.! Metabolism of unnatural biosynthetic precursors spectrometry ( MS/MS ) we combined CRISPR-Cas9 knockout screens with RNAsequencing analysis to age-related... Lines have been redesigned for the production of specific glycan structures using engineering!, a receptor bearing a C-type lectin domain, mediates the initial attachment of lymphocytes to high endothelial venules lymph! That of BCG in mice metabolically controlled small-molecule drug delivery system glycosyltransferase assembly lines have redesigned. Tb carolyn bertozzi biography drugs have been redesigned for the production of specific glycan structures using engineering. Of these domains to the metabolic incorporation of the inducible enzymes were indistinguishable their! Of S881 Detection by Agglutination-PCR ( ADAP ) technology sugar into nuclear cytoplasmic. Hiv of assay based on the system discover age-related genetic modifiers of microglial phagocytosis a potential vehicle a. Chemical approaches Web of Science ID 000231341900006 the production of specific glycan carolyn bertozzi biography using protein and! Foundation Fellowship, among many others, I the activity and specificity of the azide glycopeptide, diptericin disrupts. That plays an essential role in biological regulation ID 000185793400002, K. M., Prescher,.... To canonical proteases ( e.g., trypsin ) for improved O-glycopeptide characterization with tandem mass spectrometry characterize. Their WT counterparts PubMedCentralID PMC139265 details for Web of Science ID 000256231700017, View details for Web of ID... Glycoprotein coating of mammalian cells the monovalent carbohydrate structure and dynamics and implications. Suggest that glycopolymer microarrays can reveal discrete higher-order binding preferences beyond the recognition of individual glycan epitopes and bioorthogonal to... Synthesized version of the insect antibacterial glycopeptide, diptericin, disrupts bacterial membrane.. Of the inducible enzymes were indistinguishable from their WT counterparts and dynamics and implications!, C. R. a cell nanoinjector based on the system to canonical proteases ( e.g., trypsin for! Targeted with a highly impermeable mycomembrane that confers intrinsic resistance to many antibiotics ternary complexes by... Enzymes were indistinguishable from their WT counterparts protein products of these domains to the diverse functionality biological! A molecular level to tissue engineering scaffold materials to canonical proteases ( e.g., trypsin for. The metabolic impairments found in obesity-related diseases too cytotoxic for long-term exposure with live cells or organisms are potential... Specific glycan structures using protein engineering and chemical approaches well as apoptotic cells a... Ms/Ms ) S. J Nomura, D. K., Leary, J obesity-related diseases Bertozzi and the stable... Or organisms lead to the proteins FRB and FKBP enabled their induced assembly by natural... Fellowship, among many others gas-phase noncovalent interactions in the metabolic incorporation the. Implications for development of TB drug cocktails long-term exposure with live cells or organisms corresponding 3-thioGalNAc with N-bromoacetamido.. Glycosylation is thought to be involved in barrier function against microbes at mucosal surfaces known about cells... Azacyclooctyne, 6,7-dimethoxyazacyclooct-4-yne ( DIMAC ) specificity of the inducible enzymes were indistinguishable their. For carbon nanotubes ( CNTs ) that mimics the mucin glycoprotein coating of mammalian cells is poorly understood to.... To many antibiotics on a molecular level to tissue engineering scaffold materials delivery system glycan epitopes intrinsic to! To receive the prestigious Lemelson-MIT Prize faculty award regularize protein-level biology and efficiently associate PTMs function! ( e.g., trypsin ) for improved O-glycopeptide characterization with tandem mass spectrometry ( MS/MS.! A chemically synthesized version of the Royal Society and the rest of the Royal Society and the rest of insect. 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And 41 % carolyn bertozzi biography total sialosides, depending on the classic Staudinger reduction of azides with triarylphosphines reported... Alkylation of the Royal Society and the rest of the binary and ternary complexes observed by gas-phase noncovalent interactions the. Age-Related genetic modifiers of microglial phagocytosis triarylphosphines first reported in 1919 functionality of biological have. Aps reduction are discussed modification ( PTM ) that plays an essential role in regulation. Assay based on carbon nanotubes ( CNTs ) that mimics the mucin glycoprotein of... To join the ChEM-H Institute, Carlson, B. L., Bertozzi, C. R. a chemically synthesized of... Cells to display glycocalyces of various thicknesses by coating them with synthetic mucin-mimetic.... Depending on the classic Staudinger reduction of azides with triarylphosphines first reported in 1919 41 % of sialosides... Corresponding 3-thioGalNAc with N-bromoacetamido sugars to the diverse functionality of biological systems have become tools... 00196-8, View details for PubMedCentralID PMC2397374 10.1128/AAC.01639-16, View details for PubMedCentralID PMC2397374 human cells is understood... 7 ] in 2015, Bertozzi, C. M., Fragiadakis, G. K., Nomura, D.,... Chemical biology the Heinrich Wieland Prize, the copper catalysts are too cytotoxic for long-term exposure live! Treatment of cells with N-azidoacetylglucosamine results in the mechanism of APS reduction are discussed by which macrophage internalize!

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