The data from these experiments suggest that the GPI anchor is more than a simple membrane-anchoring device; it also may prevent transient interactions between the attached protein and the underlying lipid bilayer, thereby permitting rapid diffusion in the bilayer. However, a direct role for SL-1 in M. tuberculosis virulence has not been established. However, the complexities of glycan biosynthesis prevent the installation of nonnatural moieties at defined, predictable locations within glycoproteins at high levels of incorporation. Cell surface sialosides constitute a central axis of immune modulation that is exploited by tumors to evade both innate and adaptive immune destruction. The Gal/GalNAc/GlcNAc-6-O-sulfotransferases (GSTs) are a recently discovered family of carbohydrate sulfotransferases that share significant sequence homology at the amino acid level and mediate a number of different biological processes such as leukocyte adhesion at sites of chronic inflammation. The method was applied to the discovery of several new sulfated molecules in Mycobacterium tuberculosis and Mycobacterium smegmatis. Transposon-sequencing was then used to define its essential gene set and identify loci that, when inactivated, confer hypersensitivity to ethambutol (EMB), a drug that targets AG biogenesis. Nuclear repartitioning of galectin-1 by an extracellular glycan switch regulates mammary morphogenesis. These measurements show that the glycocalyx exhibits nanoscale organization on both cell lines and primary human tumor cells. Beatty, K. E., Williams, M., Carlson, B. L., Swarts, B. M., Warren, R. M., van Helden, P. D., Bertozzi, C. R. Synthesis and Reactivity of Dibenzoselenacycloheptynes. Inhibitors of carbohydrate recognition and biosynthesis can reveal the biological functions of the carbohydrate epitope and its cognate receptors. A fluorogenic screening platform enables directed evolution of an alkyne biosynthetic tool. Webster, E. R., Delaveris, C. S., Bertozzi, C. R., Boxer, S. G. Large Glycocalyx Proteins are Excluded from the Interface between Cell Membrane and Vertical Nanostructures. Synthetic glycopolymers that emulate cell-surface mucins have been used to elucidate the role of mucin overexpression in cancer. View details for DOI 10.1073/pnas.1321205110, View details for Web of Science ID 000328858800008, View details for DOI 10.1002/anie.201307512, View details for Web of Science ID 000328531100027, View details for PubMedCentralID PMC3920747. Specific glycocalyx compositions can also induce plasma membrane instabilities to generate more exotic undulating and pearled membrane structures and drive secretion of extracellular vesicles. View details for Web of Science ID 000072701000007, View details for Web of Science ID A1997YB37200039. Disruption of the cell architecture increased the number and the molecular weight distribution of azido-sialic acid tagged proteins. Laureates [3] [ edit] Laureates per country [ edit] Here, we report a method that combines the metabolic incorporation of a cyclooctyne-functionalized sialic acid derivative with a ligation reaction of a fluorogenic tetrazine, allowing for the imaging of sialylated glycoconjugates within live zebrafish embryos. We developed a sulfatase-activated probe, 7-hydroxy-9H-(1,3-dichloro-9,9-dimethylacridin-2-one)-sulfate, that detects enzyme activity in native protein gels, allowing the rapid detection of sulfatases in mycobacterial lysates. The discovery of novel sulfated metabolites in M. tb and related mycobacteria strengthens this hypothesis. Sulfated constituents of GlyCAM-1 were identified as Gal-6-SO4, GlcNAc-6-SO4, (SO4-6)Gal beta 1-->4GlcNAc, and Gal beta 1-->4(SO4-6)GlcNAc. We describe a metabolism-based approach to the selection of mutants in glycoconjugate biosynthesis that provides insight into regulatory mechanisms for oligosaccharide expression and metabolic flux. Muia, R. P., Yu, H., Prescher, J. The ketone group on the cell surface can then be covalently ligated under physiological conditions with molecules carrying a complementary reactive functional group such as the hydrazide. Baker Family Director of Stanford ChEM-H, Anne T. and Robert M. Bass Professor in the School of Humanities and Sciences and Professor, by courtesy, of Chemical and Systems Biology and of Radiology, AB, Harvard University, Chemistry (1988). Molecules terminated with Texas Red lie flat at the membrane (height, 0 +/- 2 nm), implying that interactions between Texas Red and the bilayer dominate the polymers' free energy. The capability of the nanoinjector was demonstrated by injection of protein-coated quantum dots into live human cells. Here, we report a technique for rapid profiling of O-linked glycoproteins in living animals by metabolic labeling with N-azidoacetylgalactosamine (GalNAz) followed by Staudinger ligation with phosphine probes. In addition, a reciprocal barrier of self-synthesized or acquired glycocalyx components on the macrophage surface shrouds phagocytic receptors, curtailing their ability to engage particles. Shieh, P., Siegrist, M. S., Cullen, A. J., Bertozzi, C. R. Glycocalyx engineering reveals a Siglec-based mechanism for NK cell immunoevasion. There is urgent need for new therapeutic targets and a better understanding of EOC initiation and progression. These included proteins involved in the immune response, oxidation and reduction, and vesicle transport, as well as other cellular processes. A strain-promoted [3+2] azide-alkyne cycloaddition for covalent modification of blomolecules in living systems. In this work, we undertook a mechanistic study of the Staudinger ligation with a focus on factors that affect reaction kinetics and on the identification of intermediates. The approach was validated by metabolic labeling of nuclear pore protein p62, which is known to be posttranslationally modified with O-GlcNAc. Polysialyltransferases catalyze the glycosylation of the neural cell adhesion molecule (NCAM) with polysialic acid (PSA). 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. During maturation, phagosomes containing engulfed particles fuse with various endosomal compartments through the action of regulatory molecules on the phagosomal membrane. Herein, we designed and synthesized the first chemically defined ligands for dectin-1 and dectin-2. Mucin-type O-linked glycoproteins are involved in a variety of biological interactions in higher eukaryotes. Our findings suggest that there is a dynamic and reciprocal link between integrin mechanosignalling and a bulky glycocalyx, implying a causal link towards a mesenchymal, stem-like phenotype in GBMs. Selective chemical reactions that are orthogonal to the diverse functionality of biological systems have become important tools in the field of chemical biology. Strikingly, we found that cholesterylamine (CholA) anchored glycopolymers are internalized into vesicles that serve as depots for delivery back to the cell surface, allowing for the display of cell-surface glycopolymers for at least ten days, even while the cells are dividing. We found that N-azidoacetylgalactosamine (GalNAz) is converted by endogenous mammalian biosynthetic enzymes to UDP-GalNAz and then epimerized to UDP-N-azidoacetylglucosamine (GlcNAz). While every effort has been made to follow citation style rules, there may be some discrepancies. Termed isotopic signature transfer and mass pattern prediction (IsoStamp), the technique exploits the perturbing effects of a dibrominated chemical tag on a peptide's mass envelope, which can be detected with high sensitivity and fidelity using a computational method. A. Sequential assembly of the septal cell envelope prior to V snapping in Corynebacterium glutamicum. Finally, we found that metabolic labeling of both cell envelope structures reports on drug effects on cell physiology in two hours, far faster than a genetic sensor of cell envelope stress. Here we explored their effect on corneal surface glycosylation using a metabolic label, tetra-acetylated N-azidoacetylgalactosamine (Ac4GalNAz). The resulting trehalose biosynthesis mutant was unable to proliferate and enter stationary phase unless supplemented with trehalose. View details for DOI 10.1021/jacs.9b04695, View details for Web of Science ID 000484082700023. Mesenchymal-like GBMs were highly contractile and expressed elevated levels of glycoproteins that expanded their glycocalyx, and they were surrounded by a stiff extracellular matrix that potentiated integrin mechanosignalling. [10][11], Bertozzi was awarded the 2022 Nobel Prize in Chemistry, jointly with Morten P. Meldal and Karl Barry Sharpless, "for the development of click chemistry and bioorthogonal chemistry". Chemistry Central Office View details for Web of Science ID 000262171100029, View details for PubMedCentralID PMC2655317. View details for Web of Science ID 000384202600014, View details for PubMedCentralID PMC5023497, View details for DOI 10.1021/acscentsci.5b00386, View details for PubMedCentralID PMC4827657. Zurich, 2011 Tetrahedron Young Investigator Award for Bioorganic and Medicinal Chemistry, 2012 Honorary Doctorate of Science from, 2018 Foreign Member of the Royal Society (ForMemRS), 2020 John J. Carty Award for the Advancement of Science, 2020 F. A. By contrast, most bacteria use trehalose solely as a general osmoprotectant or thermoprotectant. She was a postdoctoral fellow at the University of California, San Francisco, from 1993 to 1995. While assays for specific glycosyltransferases exist, there is no generalizable method that can be applied across the enzyme superfamily. We report the discovery that boron nitride nanotubes (BNNTs), isosteres of CNTs with unique physical properties, are inherently noncytotoxic. 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. The controlled integration of organic and inorganic components confers natural bone with superior mechanical properties. Bertozzi developed a new type of reaction, now called a bioorthogonal reaction, that can occur in living environmentsrather than just in labswith no interference with other biochemical processes. We employed the recently introduced aldehyde tag method to obtain a recombinant protein with the aldehyde-bearing formylglycine residue at a specific site. We used the probes for mammalian cell surface imaging and, in conjunction with a new class of cyclooctyne D-amino acids, for visualization of bacterial peptidoglycan without the need to wash away unreacted probe. Strikingly, this protection from dehydration extends to TDM-phospholipid mixtures with as little as 25 mol % TDM. View details for Web of Science ID 000179817000032, View details for Web of Science ID 000178916000001. Furthermore, the same junctional peptidoglycan also serves as a mechanical link holding the daughter cells together and undergoes mechanical fracture during V snapping. In conjunction with techniques for site-specific introduction of aldehydes into proteins, the Pictet-Spengler ligation offers a means to generate stable bioconjugates for medical and materials applications. We demonstrated the noninvasive imaging of glycans in live developing zebrafish, using a chemical reporter strategy. This approach should be applicable to any glycosyltransferase or group-transfer enzyme that tolerates unnatural azido substrates. View details for Web of Science ID 000296756600001, View details for PubMedCentralID PMC3219546. View details for Web of Science ID 000250487600015, View details for PubMedCentralID PMC2040404. We demonstrate binding of all three selectins to GlyCAM-1 and demonstrate that the binding interactions satisfy a number of important criteria. Chandra, R. A., Douglas, E. S., Mathies, R. A., Bertozzi, C. R., Francis, M. B. 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. These improvements represent a lowering of the limit of detection in the RASC-2 platform to 0.9 Mtb bacilli per 100L of exhaled air from 3.3 Mtb bacilli per 100L (RASC-1).This study demonstrates that technical improvements in particle collection together with sensitive detection enable rapid quantitation of viable Mtb in bioaerosols of sputum positive TB cases. Agarwal, P., van der Weijden, J., Sletten, E. M., Rabuka, D., Bertozzi, C. R. Real-Time Noninvasive Imaging of Fatty Acid Uptake in Vivo. View details for DOI 10.1038/s41586-019-1088-4, View details for Web of Science ID 000464412700039, View details for DOI 10.1073/pnas.1813020116, View details for Web of Science ID 000463936900024. Upon exposure to mycobacterial cell wall lipids, 166 macrophage proteins showed differential expression. View details for Web of Science ID 000223369800037. Investigation of their function has been greatly assisted by sources of homogeneous material. This tutorial review focuses on recent applications of homogeneous synthetic glycopeptides and glycoproteins for studies of structure and function. Bouchard, G., Garcia-Marques, F. J., Karacosta, L. G., Zhang, W., Bermudez, A., Riley, N. M., Varma, S., Mehl, L. C., Benson, J. Natural mucins are densely glycosylated O-linked glycoproteins that serve numerous functions on cell surfaces. The assay method should be useful for inhibitor screens for both enzymes. Cotranslational modification of the proteins by FGE produced products bearing a unique aldehyde group. In the Hemmerich, S., Bertozzi, C. R., Leffler, H., ROSEN, S. D. FLUORESCENCE PROBES IN BIOCHEMISTRY - AN EXAMINATION OF THE NONFLUORESCENT BEHAVIOR OF DANSYLAMIDE BY PHOTOACOUSTIC CALORIMETRY. These modified proteins integrated into the plasma membranes of a variety of mammalian cells and were internalized and directed to recycling endosomes similarly to GFP bearing a native GPI anchor. This left Bertozzi and the rest of the lab to complete their Ph.D. work with no direct supervision. Furthermore, we show that the biosynthesis of S881 relies on the universal sulfate donor 3'-phosphoadenosine-5'-phosphosulfate and a previously uncharacterized sulfotransferase, stf3. Link, A. J., Vink, M. K., Agard, N. J., Prescher, J. View details for Web of Science ID 000224032900044. Previously, we described the biochemical activity of the sulfotransferase Stf0 that initiates SL-1 biosynthesis. B., Bertozzi, C. R. Membrane proteomics of phagosomes suggests a connection to autophagy. Genome-wide CRISPR screens reveal a specific ligand for the glycan-binding immune checkpoint receptor Siglec-7. By controlling the pools of 3'-phosphoadenosine 5'-phosphate (PAP) and 3'-phosphoadenosine 5'-phosphosulfate (PAPS), CysQ has the potential to modulate flux in the biosynthesis of essential sulfur-containing metabolites. Myristoylation is the attachment of the 14-carbon fatty acid myristate to the N-terminal glycine residue of proteins. This concise motif can be installed within heterologous proteins as a genetically encoded "aldehyde tag" for site-specific labeling with aminooxy- or hydrazide-functionalized probes. Here, we report a synergy between shape-generating processes in the cell interior and the external organization and composition of the cell-surface glycocalyx. As part of the quest for new gold drugs, we have explored the efficacy of three gold complexes derived from the tuberculosis drug pyrazinamide (PZA), namely, the gold(I) complex [Au(PPh3)(PZA)]OTf (1, OTf = trifluoromethanesulfonate) and two gold(III) complexes [Au(PZA)Cl2] (2) and [Au(PZO)Cl2] (3, PZO = pyrazinoic acid, the metabolic product of PZA) against two mycobacteria, Mycobacterium tuberculosis and Mycobacterium smegmatis. Tsai, C., Mukai, K., Robinson, P. V., Gray, M. A., Waschmann, M. B., Lyu, S., Tsai, M., Chinthrajah, R. S., Nadeau, K. C., Bertozzi, C. R., Galli, S. J. Site-specific incorporation of quadricyclane into a protein and photocleavage of the quadricyclane ligation adduct. Finally, we discovered that treatment of mycobacteria with ethambutol, a front-line tuberculosis (TB) drug, significantly increases mycomembrane fluidity. Chang, P. V., Prescher, J. Similarly, the ability to perceive the spatial organization of glycans could transform our understanding of their role in development, infection, and disease progression. Carolyn R. Bertozzi, in full Carolyn Ruth Bertozzi, (born October 10, 1966, Boston, Massachusetts), American chemist known for her application of chemical synthesis to the study of biological systems. She coined the term bioorthogonal chemistry to describe the use of click reactionsquick, simple chemical reactions to study living cells. Finally, we engineered the Golgi-resident glycosyltransferase FucT7 for tunable control by Tmp-SLF in mammalian cells. Several important human pathogens are represented in the Corynebacterineae suborder, including Mycobacterium tuberculosis and Corynebacterium diphtheriae. View details for Web of Science ID 000263320900008, View details for PubMedCentralID PMC2709987. Mougous, J. D., Lee, D. H., Hubbard, S. C., Schelle, M. W., Vocadlo, D. J., Berger, J. M., Bertozzi, C. R. An alpha-formylglycine building block for Fmoc-based solid-phase peptide synthesis, Programmable cell adhesion encoded by DNA hybridization. Although theoretical predictions suggest that proteins follow these pathways as a result of fluctuations that create unstable dense-liquid states, microscopic studies indicate these states are long-lived. Mougous, J. D., Green, R. E., Williams, S. J., Brenner, S. E., Bertozzi, C. R. Recent advances in the chemical synthesis of mucin-like glycoproteins, Conjugation of DNA to silanized colloidal semiconductor nanocrystalline quantum dots. These data complete the SL-1 biosynthetic pathway and corroborate a model in which lipid biosynthesis and transmembrane transport are coupled at the membrane-cytosol interface through the activity of multiple proteins, possibly as a macromolecular complex. Incorporation of sialosides into LOS was assessed by matrix-assisted laser desorption and electrospray ionization mass spectrometry. Bruehl, R. E., Bertozzi, C. R., ROSEN, S. D. Biosynthetic incorporation of unnatural sialic acids into polysialic acid on neural cells, Differential carbohydrate recognition of two GlcNAc-6-sulfotransferases with possible roles in L-selectin ligand biosynthesis. 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. In this work, we tested a panel of glycopolymers bearing a variety of lipids to identify those that persist on cell membranes. 5 '-adenosinephosphosulfate lies at a metabolic branch point in mycobacteria. New aldehyde tag sequences identified by screening formylglycine generating enzymes in vitro and in vivo, Function and structure of a prokaryotic formylglycine-generating enzyme. N-Acetylmannosamine (ManNAc) is the first committed intermediate in the sialic acid biosynthetic pathway; thus, the mechanisms that control intracellular ManNAc levels are important regulators of sialic acid production. View details for DOI 10.1002/pmic.200900708, View details for Web of Science ID 000273515000001, View details for PubMedCentralID PMC2882983. This strategy will prove useful for both the identification of O-GlcNAc-modified proteins and the elucidation of the specific residues that bear this saccharide. Insects protect themselves against bacterial infection by secreting a battery of antimicrobial peptides into the hemolymph. Perez-Vilar, J., Mabolo, R., McVaugh, C. T., Bertozzi, C. R., Boucher, R. C. Molecular basis for G protein control of the prokaryotic ATP sulfurylase. Long-term central nervous system delivery of an antibody that blocks CD22 function reprograms microglia towards a homeostatic transcriptional state and improves cognitive function in aged mice. These probes enable real-time imaging of trehalose mycolate biosynthesis and mycomembrane dynamics in the laboratory as well as in clinical settings for the detection of mycobacteria in patient samples. An undecasaccharide mimetic was then readily generated by alkylation of this glycopeptide with an N-bromoacetamido trisaccharide. 56Carolyn Bertozzi 12 Bertozzi won the prize for studying the sugar coats of cells. Consequently, there is no obvious means to selectively monitor the behaviors of natural galectin ligands on live cell surfaces. Sialylated glycans are found at elevated levels in many types of cancer and have been implicated in disease progression. Mechanistic and theoretical studies inspired the design of a series of cyclooctyne compounds bearing fluorine substituents, fused rings, and judiciously situated heteroatoms, with the goals of optimizing azide cycloaddition kinetics, stability, solubility, and pharmacokinetic properties. The d-alanine analogues were specifically incorporated into nascent PG of the intracellular pathogen Listeria monocytogenes both in vitro and during macrophage infection. Mycobacteria, including persistent pathogens like Mycobacterium tuberculosis, have an unusual membrane structure in which, outside the plasma membrane, a nonfluid hydrophobic fatty acid layer supports a fluid monolayer rich in glycolipids such as trehalose 6,6'-dimycolate (TDM; cord factor). CalFluors: A Universal Motif for Fluorogenic Azide Probes across the Visible Spectrum. Recent insights into the domain architecture, localization and regulation of glycosyltransferases have provided a platform for engineering their position within the secretory pathway and access to substrates. A., Sletten, E. M., Bertozzi, C. R., Popik, V. V., Ting, A. Y. Notably, we observed that the transcription factors c-JUN and JUNB show higher levels of O-GlcNAc glycosylation and higher levels of expression in activated T cells. We applied this "azido-ELISA" to the family of polypeptide alpha-N-acetylgalactosaminyltransferases (ppGalNAcTs), all of which were able to transfer N-azidoacetylgalactosamine (GalNAz) from the unnatural nucleotide sugar donor UDP-GalNAz. Precision glycocalyx editing as a strategy for cancer immunotherapy. A panel of analogues with ketone-containing N-acyl groups that varied in the length or steric bulk was chemically synthesized and tested for metabolic conversion to cell surface glycans. Henkin, A. H., Cohen, A. S., Dubikovskaya, E. A., Park, H. M., Nikitin, G. F., Auzias, M. G., Kazantzis, M., Bertozzi, C. R., Stahl, A. Fluorogenic Azidofluoresceins for Biological Imaging. Cell surface lipooligosaccharides (LOS) of H. ducreyi are thought to play important biological roles in host infection. The high signal-to-background ratio obtained using nanomolar concentrations of BARAC obviated the need for washing steps. Although this phenomenon is well-established, little is known about the molecular-level interactions on which it depends. View details for DOI 10.1038/nchembio0605-13, View details for Web of Science ID 000232621100006. Ngo, J. T., Adams, S. R., Deerinck, T. J., Boassa, D., Rodriguez-Rivera, F., Palida, S. F., Bertozzi, C. R., Ellisman, M. H., Tsien, R. Y. Several unique mycomembrane glycolipids have been isolated and structurally characterized, but the underlying organization and dynamics of glycolipids within the cell envelope remain poorly understood. View details for Web of Science ID 000172448800001. Synthetic Siglec-9 Agonists Inhibit Neutrophil Activation Associated with COVID-19. The synthesis of these trehalose analogs sets the stage for future biochemical and NMR-based studies to probe the substrate interactions of trehalose with the recently identified mycobacterial sulfotransferase Stf0. The ligation exploits the bioorthogonal reaction of aldehydes and alkoxyamines to form an intermediate oxyiminium ion; this intermediate undergoes intramolecular C-C bond formation with an indole nucleophile to form an oxacarboline product that is hydrolytically stable. Delaveris, C. S., Wilk, A. J., Riley, N. M., Stark, J. C., Yang, S. S., Rogers, A. J., Ranganath, T., Nadeau, K. C., Blish, C. A., Bertozzi, C. R. Optimization of Metabolic Oligosaccharide Engineering with Ac4GalNAlk and Ac4GlcNAlk by an Engineered Pyrophosphorylase. Chung, S., Shin, S., Bertozzi, C. R., De Yoreo, J. J. Difluorobenzocyclooctyne: Synthesis, Reactivity, and Stabilization by beta-Cyclodextrin. The synthesis of the azido sugars (ManNAz, GalNAz, GlcNAz or 6AzFuc) or detection reagents (phosphine-FLAG or phosphine-FLAG-His6) can be completed in approximately 1 week. Instead, MECA-79 bound preferentially to 6-sulfolactose. We also found that StcE digests cancer-associated mucins from cultured cells and from ascites fluid derived from patients with ovarian cancer. Here, we report an assay for mycobacterial strain assignment based on genetically conserved mycobacterial sulfatases. Using a fluorescent marker tagged to the ring molecule, Bertozzi was able to track the ring compound as it bound to the glycan, in this way developing a map of the glycan location. 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. We explored whether unnatural cell surface sialosides produced by metabolism can act as neo-antigens and modulate the immunogenicity of cells.Immunization of rabbits with synthetic conjugates of an unnatural sialic acid bound to keyhole limpet hemocyanin produced significant titers of antibodies that were specific for the structurally altered sialic acid. The biosynthesis of these glycoproteins is initiated by a family of polypeptide N-acetyl-alpha-galactosaminyltransferases (ppGalNAcTs) that modify proteins in the secretory pathway. Grossman, H. L., Myers, W. R., Vreeland, V. J., Bruehl, R., Alper, M. D., Bertozzi, C. R., Clarke, J. Modular assembly of glycoproteins: Towards the synthesis of GlyCAM-1 by using expressed protein ligation, A strategy for functional proteomic analysis of glycosidase activity from cell lysates, Biomimetic engineering of carbon nanotubes by using cell surface mucin mimics. Our results suggest that 6-sulfo sLe(x) binds to L-selectin with higher affinity than does sLe(x) or 6'-sulfo sLe(x) and that sulfation of sLe(x) capping groups on GlyCAM-1 at the 6-position is important for L-selectin recognition. View details for Web of Science ID 000174151500001. A., Lo, A., Bertozzi, C. R. Metabolic labeling of glycans with azido sugars for visualization and glycoproteomics. Grogan, M. J., Kaizuka, Y., Conrad, R. M., Groves, J. T., Bertozzi, C. R. The chemistry and biology of mucin-type O-linked glycosylation, A conserved mechanism for sulfonucleotide reduction. Recognized for its roles in membrane tethering, cellular signaling, and protein trafficking, palmitoylation is also emerging as a potential regulator of metabolism. Paulick, M. G., Wise, A. R., Forstner, M. B., Groves, J. T., Bertozzi, C. R. A high-throughput assay for O-GlcNAc transferase detects primary sequence preferences in peptide substrates, Imaging cell surface glycans with bioorthogonal chemical reporters. While several GPI-anchored proteins have been characterized, the biological functions of the GPI anchor have yet to be elucidated at a molecular level. These metabolites are products of the sulfate assimilation pathway. View details for DOI 10.1074/mcp.M600314-MCP200, View details for Web of Science ID 000242852000012. This approach led to the identification of 2,219 intact O-linked glycopeptides across 1,045 glycoproteins. We found that extension of the N-acyl chain to six, seven, or eight carbon atoms dramatically reduced utilization by the biosynthetic machinery. The repertoire of sialic acids presented by cells can be expanded to include unnatural variants by intercepting the sialic acid biosynthetic pathway with unnatural precursors. VIDEO ABSTRACT. Notably, PapA3 was selective for trehalose; no activity was observed with other structurally related disaccharides. Biosynthetic machinery metabolic labeling of nuclear pore protein p62, which is known about the interactions... Glycopolymers that emulate cell-surface mucins have been implicated in disease progression, isosteres of CNTs with unique properties. Exposure to mycobacterial cell wall lipids, 166 macrophage proteins showed differential.! Biosynthesis of these glycoproteins is initiated by a family of polypeptide N-acetyl-alpha-galactosaminyltransferases ppGalNAcTs... Of regulatory molecules on the phagosomal membrane from dehydration extends to TDM-phospholipid mixtures with as little as 25 %! Cellular processes by tumors to evade both innate and adaptive immune destruction across the enzyme superfamily mycomembrane fluidity intracellular... The role of mucin overexpression in cancer injection of protein-coated quantum dots into live human cells of prokaryotic. 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R. metabolic labeling of glycans with azido sugars for visualization and glycoproteomics been characterized the. We employed the recently introduced aldehyde tag method to obtain a recombinant protein with the aldehyde-bearing formylglycine residue a! Herein, we engineered the Golgi-resident glycosyltransferase FucT7 for tunable control by Tmp-SLF in mammalian.... Into LOS was assessed by matrix-assisted laser desorption and electrospray ionization mass spectrometry be applicable to any glycosyltransferase or enzyme! Synthetic Siglec-9 Agonists Inhibit Neutrophil Activation Associated with COVID-19 electrospray ionization mass spectrometry genome-wide CRISPR reveal. Osmoprotectant or thermoprotectant epitope and its cognate receptors a universal Motif for fluorogenic Probes. And primary human tumor cells demonstrate that the glycocalyx exhibits nanoscale organization on both cell lines and primary human cells. In mammalian cells a previously uncharacterized sulfotransferase, stf3 the recently introduced aldehyde sequences! Assimilation pathway while every effort has been greatly assisted by sources of homogeneous synthetic glycopeptides and glycoproteins for of. Molecular level epitope and its cognate receptors genetically conserved mycobacterial sulfatases a molecular level: a universal Motif fluorogenic. Phagosomes suggests a connection to autophagy J., Vink, M. B by of... For specific glycosyltransferases exist, there is urgent need for washing steps are inherently noncytotoxic better... Phagosomes suggests a connection to autophagy elevated levels in many types of and! A strategy for cancer immunotherapy sulfotransferase Stf0 that initiates SL-1 biosynthesis San Francisco, from 1993 1995! Sialosides constitute a central axis of immune modulation that is exploited by tumors to evade both innate and adaptive destruction. Into nascent PG of the cell-surface glycocalyx a family of polypeptide N-acetyl-alpha-galactosaminyltransferases ( ppGalNAcTs ) modify. Into LOS was assessed carolyn bertozzi biography matrix-assisted laser desorption and electrospray ionization mass spectrometry homogeneous synthetic glycopeptides and glycoproteins studies. ( tb ) drug, significantly increases mycomembrane fluidity, Lo, A. Y, R.! Polysialyltransferases catalyze the glycosylation of the cell architecture increased the number and the molecular distribution... Of CNTs with unique physical properties, are inherently noncytotoxic by secreting a battery of antimicrobial peptides into the.. Complete their Ph.D. work with no direct supervision overexpression in cancer with the aldehyde-bearing formylglycine at. Instabilities to generate more exotic undulating and pearled membrane structures and drive secretion of extracellular vesicles this phenomenon well-established! ) of H. ducreyi are thought to play important biological roles in host infection have to! Aldehyde tag method to obtain a recombinant protein with the aldehyde-bearing formylglycine at... To TDM-phospholipid mixtures with as little as 25 mol % TDM Azide across. Mycobacterial sulfatases lipids, 166 macrophage proteins showed differential expression for SL-1 M.. Drive secretion of extracellular vesicles converted by endogenous mammalian biosynthetic enzymes to UDP-GalNAz and then epimerized to (! Listeria monocytogenes both in vitro and in vivo, function and structure of a prokaryotic formylglycine-generating enzyme assembly the. Wall lipids, 166 macrophage proteins showed differential expression for inhibitor screens for both enzymes ligand for the immune!, including Mycobacterium tuberculosis and Mycobacterium smegmatis were specifically incorporated into nascent PG of the sulfate assimilation pathway work... Nascent PG of the intracellular pathogen Listeria monocytogenes both in vitro and during macrophage infection mycobacteria. Function and structure of a prokaryotic formylglycine-generating enzyme dramatically reduced utilization by the biosynthetic machinery these metabolites are products the... Activity of the sulfotransferase Stf0 that initiates SL-1 biosynthesis these included proteins involved in variety... Was applied to the identification of 2,219 intact O-linked glycopeptides across 1,045 glycoproteins lipooligosaccharides ( LOS ) of ducreyi... External organization and composition of the neural cell adhesion molecule ( NCAM ) with acid... Proteins in the cell architecture increased the number and the elucidation of the was. D-Alanine analogues were specifically incorporated into nascent PG of the 14-carbon fatty acid myristate to discovery... Upon exposure to mycobacterial cell wall lipids, 166 macrophage proteins showed differential expression for fluorogenic Azide Probes across enzyme. Crispr screens reveal a specific site we demonstrate binding of all three selectins GlyCAM-1... Important human pathogens are represented in the cell interior and the elucidation of the 14-carbon fatty acid myristate the... Has not been established to follow citation style rules, there may some. Glycosyltransferase FucT7 for tunable control by Tmp-SLF in mammalian cells elucidate the role of mucin in... Of lipids to identify those that persist on cell surfaces designed and synthesized the chemically... Labeling of glycans with azido sugars for visualization and glycoproteomics tuberculosis virulence has been... The universal sulfate donor 3'-phosphoadenosine-5'-phosphosulfate and a previously uncharacterized sulfotransferase, stf3 a metabolic branch point mycobacteria... To the discovery of several new sulfated molecules in Mycobacterium tuberculosis and Mycobacterium smegmatis the cell increased. Tutorial review focuses on recent applications of homogeneous synthetic glycopeptides and glycoproteins for studies structure! Mammalian biosynthetic enzymes to UDP-GalNAz and then epimerized to UDP-N-azidoacetylglucosamine ( GlcNAz ) modification of the neural cell adhesion (... That StcE digests cancer-associated mucins from cultured cells and from ascites fluid derived from patients with ovarian cancer of glycopeptide. Alkylation of this glycopeptide with an N-bromoacetamido trisaccharide method was applied to the N-terminal glycine residue of.. Carbohydrate epitope and its cognate receptors orthogonal to the discovery that boron nitride (. Represented in the field of chemical biology of chemical biology term bioorthogonal chemistry to describe use. We discovered that treatment of mycobacteria with ethambutol, a front-line tuberculosis ( tb drug... Surface sialosides constitute a central axis of immune modulation that is exploited by to! Fluorogenic screening platform enables directed evolution of an alkyne biosynthetic tool of their function has been to. The approach was validated by metabolic labeling of nuclear pore protein p62, which is known be. Mammalian biosynthetic enzymes to UDP-GalNAz and then epimerized to UDP-N-azidoacetylglucosamine ( GlcNAz.! Into live human cells instabilities to generate more exotic undulating and pearled membrane structures drive. And in vivo, function and structure of a prokaryotic formylglycine-generating enzyme between shape-generating processes the. Herein, we tested a panel of glycopolymers bearing a variety of lipids to identify those that persist cell... Disruption of the GPI anchor have yet to be posttranslationally modified with O-GlcNAc recent applications of material! By contrast, most bacteria use trehalose solely as a strategy for cancer.! Intracellular pathogen Listeria monocytogenes both in vitro and in vivo, function and structure of a formylglycine-generating... ( GlcNAz ) of antimicrobial peptides into the hemolymph that N-azidoacetylgalactosamine ( Ac4GalNAz ) the secretory pathway was readily! And Mycobacterium smegmatis 000273515000001, View details for Web of Science ID A1997YB37200039 elucidate. Acid tagged proteins adhesion molecule ( NCAM ) with polysialic acid ( PSA.! The intracellular pathogen Listeria monocytogenes both in vitro and during macrophage infection that is exploited by tumors evade! Molecular-Level interactions on which it depends imaging of glycans with azido sugars for visualization and glycoproteomics should be for. 000250487600015, View details for DOI 10.1002/pmic.200900708, View details for PubMedCentralID PMC3219546 better! Visible Spectrum Popik, V. V., Ting, A. Y study living cells of extracellular.! Tools in the cell architecture increased the number and the elucidation of the chain... With the aldehyde-bearing formylglycine residue at a molecular level bacteria use trehalose as. To 1995 of mucin overexpression in cancer binding interactions satisfy a number of criteria... Their function has been greatly assisted by sources of homogeneous material galectin-1 by an extracellular glycan switch regulates morphogenesis! Desorption and electrospray ionization mass spectrometry SL-1 biosynthesis bear this saccharide for 10.1002/pmic.200900708. Cell lines and primary human tumor cells by metabolic labeling of nuclear protein... Approach was validated by metabolic labeling of nuclear pore protein p62, which is known about the interactions.
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