The data presented here was obtained with the application of a bioorthogonal chemical reporter strategy analyzing differential glycoprotein expression following the knock-down (KD) of the GALNT3 gene in the epithelial ovarian cancer (EOC) cell line A2780s. The glycosylphosphatidylinositol (GPI) anchor is a C-terminal posttranslational modification found on many eukaryotic proteins that reside in the outer leaflet of the cell membrane. Ooi, Y. S., Majzoub, K., Flynn, R. A., Mata, M. A., Diep, J., Li, J. K., van Buuren, N., Rumachik, N., Johnson, A. G., Puschnik, A. S., Marceau, C. D., Mlera, L., Grabowski, J. M., Kirkegaard, K., Bloom, M. E., Sarnow, P., Bertozzi, C. R., Carette, J. E. A novel therapeutic modality of inhibiting the glyco-immune checkpoint axis to treat cancer. Each enzyme preferred a terminal GlcNAc residue, and was impeded by the addition of a beta1,4-linked Gal residue (i.e., terminal LacNAc). StcEE447D is a pan-mucin stain derived from enterohemorrhagic Escherichia coli that is tolerant to a wide range of glycoforms. Her biography is available in 16 different languages on Wikipedia. Three piperidine-melamine glycodendrimers were synthesized via a chemoenzymatic route and attached to human growth hormone and the Fc region of human IgG. Nijmegen) (2013); Elected Fellow of the National Academy of Inventors (2013); Heinrich Wieland Prize (2012); Elected member of the Institute of Medicine (2011); Tetrahedron Young Investigator Award for Bioorganic and Medicinal Chemistry (2011); Royal Society of Chemistry - Organic Division, Bioorganic Chemistry Award (2010); Lemelson-MIT Prize (2010); Albert Hofmann Medal, Univ. These multivalent arrays are 4 orders of magnitude better than the monovalent carbohydrate. The GlcNAc-6-sulfotransferases are a family of Golgi-resident enzymes that modulate glycan function. A combination of PZA and the {Au(PPh3)}-thiol polymeric species appears to lead to enhanced efficacy of 1 against M. tuberculosis. It is anticipated that the ability of this technique to create virtually any type of 2D heterogeneous cell pattern should prove highly useful for the examination of key questions in cell signaling, including stem cell differentiation and cancer metastasis. These uridine analogs represent the first generation of chemical tools to study the functions of mucin-type O-linked glycosylation. Using this new method, the glycome analysis of cell membranes isolated from human embryonic stem cells and somatic cell lines was performed. Previously, we reported targeting of the blue fluorophore coumarin to cellular proteins fused to a 13-amino acid recognition sequence (LAP), catalyzed by a mutant of the Escherichia coli enzyme lipoic acid ligase (LplA). We show that Glyco-seek detects attomoles of glycoproteins of interest from cell lysates, with sensitivity several orders of magnitude higher than that of current techniques. WebCarolyn Bertozzi (1966-ngin 10-ngiet 10-ngit ) he M-koet ke yit-chak fa-hok-k. 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. [40] It focuses on biotechnologies for at-home diagnoses for type 1 diabetes, HIV, and other diseases. During the past two years, significant progress has been made in the design and synthesis of carbohydrate-based inhibitors of selectins, receptors involved in the attachment of leukocytes to endothelial cells at sites of inflammation. 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. The Leu-13 --> Gly (L13G) mutation is found in each of the three MetRS mutants, and the MetRS variant containing this single mutation is highly efficient in producing recombinant proteins that contain azidonorleucine. 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. Rodriguez-Rivera, F. P., Zhou, X., Theriot, J. View details for Web of Science ID 000257257100003, View details for PubMedCentralID PMC2663890. 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. The development of chemical strategies for decorating cells with defined carbohydrate epitopes would greatly facilitate studies of carbohydrate-mediated cell surface interactions. We introduce a non-natural substrate biosynthetic pathway and use engineered glycosyltransferases to incorporate chemically tagged sugars into the cell surface glycome of the living cell. The dependence of the desiccation protection on the synthetic trehalose glycolipid fraction is nearly identical to that of TDM. View details for DOI 10.1002/anie.201508861, View details for Web of Science ID 000368061800024, View details for PubMedCentralID PMC4715665. 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. A. Inverting family GH156 sialidases define an unusual catalytic motif for glycosidase action. WebCarolyn Ruth Bertozzi (born October 10, 1966) is an American chemist and Nobel laureate, known for her wide-ranging work spanning both chemistry and biology. The concept of a folding funnel with kinetic traps describes folding of individual proteins. Fernandez-Suarez, M., Baruah, H., Martinez-Hernandez, L., Xie, K. T., Baskin, J. M., Bertozzi, C. R., Ting, A. Y. Molecular orientation of membrane-anchored mucin glycoprotein mimics. The assay is based on the release of tritiated formaldehyde from UDP-galactofuranose but not UDP-galactopyranose by periodate and was used to identify a uridine-based enzyme inhibitor from a chemical library. View details for DOI 10.1073/pnas.1012864107, View details for Web of Science ID 000285521500020, View details for PubMedCentralID PMC3003011. View details for DOI 10.1074/mcp.M600314-MCP200, View details for Web of Science ID 000242852000012. 2023 Chuan He Hiroaki Suga Jeffery W. Kelly The sialome comprises sialylated glycoproteins and glycolipids that play essential roles in cell-cell communication. Cell surface reactions of this kind should prove useful in the introduction of new recognition epitopes, such as peptides, oligosaccharides, or small organic molecules, onto cell surfaces and in the subsequent modulation of cell-cell or cell-small molecule binding events. Alternatively, selective inhibition or activation of glycosyltransferases or glycosidases can define the biological roles of the corresponding glycans. Laughlin, S. T., Baskin, J. M., Amacher, S. L., Bertozzi, C. R. Rapid detection, discovery, and identification of post-translationally myristoylated proteins during apoptosis using a bio-orthogonal azidomyristate analog. Proteasome inhibitors are used to treat blood cancers such as multiple myeloma (MM) and mantle cell lymphoma. View details for DOI 10.1002/anie.201504249, View details for DOI 10.1021/jacs.5b04279, View details for Web of Science ID 000360321100003, View details for DOI 10.1021/acscentsci.5b00275, View details for PubMedCentralID PMC4827550. [9] Yang, A. C., Stevens, M. Y., Chen, M. B., Lee, D. P., Stahli, D., Gate, D., Contrepois, K., Chen, W., Iram, T., Zhang, L., Vest, R. T., Chaney, A., Lehallier, B., Olsson, N., du Bois, H., Hsieh, R., Cropper, H. C., Berdnik, D., Li, L., Wang, E. Y., Traber, G. M., Bertozzi, C. R., Luo, J., Snyder, M. P., Elias, J. E., Quake, S. R., James, M. L., Wyss-Coray, T. Membrane-tethered mucin-like polypeptides sterically inhibit binding and slow fusion kinetics of influenza A virus. Professor of Chemical & Systems Biology and Radiology Notably, the reaction of azides with terminal alkynes can be accelerated dramatically by copper catalysis (this highly popular Cu-catalyzed azide-alkyne cycloaddition (CuAAC) is a quintessential "click" reaction). However, because they are internalized within hours, these glycopolymers could not be employed to probe processes that occur on longer time scales. In this study we demonstrate that polysialyltransferases are capable of accepting unnatural substrates in terminally differentiated human neurons. The foundation focuses on curing NGLY1 deficiency through developing therapeutics that are efficient and inexpensive. Unnatural intermediates are used to challenge a specific pathway, and cell surface expression of their metabolic products provides a readout of flux in that pathway and a basis for selecting genetic mutants. VIDEO ABSTRACT. Upon changing to differentiation media, myotubes formed in the center of the patterned areas with an excellent degree of edge alignment. The primary sequence of APS reductase is distinguished by a conserved iron-sulfur cluster motif, -CC-X( approximately )(80)-CXXC-. One such glycolipid, Sulfolipid-1 (SL-1), consists of a trehalose-2-sulfate (T2S) core acylated with four lipid moieties. Here we show that the cytosolic enzyme N-glycanase 1 (NGLY1, the human PNGase) is essential for Nrf1 activation in response to proteasome inhibition. The kinetics of this reaction are of paramount importance for studies of dynamic processes, particularly in living subjects. Given the widespread occurrence of eukaryotic-like Ser/Thr protein kinases in bacteria, these proteins may play a broad role in bacterial osmosensing. Using high-performance liquid chromatography, we quantified the degree of accumulation and reversibility upon acidic compartment neutralization in macrophages and observed that accumulation was greater in infected than in uninfected macrophages. However, the resulting C=N linkages are susceptible to hydrolysis under physiologically relevant conditions, which limits the utility of such conjugates in biological systems. These receptors have attracted great attention in recent years due to their participation in a number of acute and chronic inflammatory diseases. However, inside infected macrophages, bacteria encounter an environment which differs substantially from broth culture and are subject to important host-dependent pharmacokinetic phenomena which modulate drug activity. Oral fluid (OF) is a highly effective substrate for population-based HIV screening efforts, as it is noninfectious and significantly easier to collect than blood. While assays for specific glycosyltransferases exist, there is no generalizable method that can be applied across the enzyme superfamily. Our study supports an essential role of one of the GalNAc-Ts - GALNT3, in EOC dissemination, including its implication in modulating PTMs and EOC metabolism. Born: 10 October 1966, Boston, MA, USA. Lee, J. H., Baker, T. J., Mahal, L. K., Zabner, J., Bertozzi, C. R., WIEMER, D. F., Welsh, M. J. Exploiting differences in sialoside expression for selective targeting of MRI contrast reagents. These data highlight the power of advanced imaging methods to provide molecular and functional insights into glycocalyx biology. View details for DOI 10.1021/acsinfecdis.6b00106, View details for Web of Science ID 000388161300007. Grogan, M. J., Pratt, M. R., Marcaurelle, L. A., Bertozzi, C. R. Kinetic parameters for small-molecule drug delivery by covalent cell surface targeting, Synthesis of oxime-linked mucin mimics containing the tumor-related T-N and sialyl T-N antigens. The prokaryotic homolog exhibits remarkable structural similarity to human FGE, including the position of catalytic cysteine residues. For example, chemical glycoproteomics technologies have enabled the identification of specific glycosylation sites and glycan structures that modulate protein function in a number of biological processes. View details for Web of Science ID 000250428200015. Enlightened by our proteomic data, we performed further experiments to show that only the LAM from M. tuberculosis inhibits accumulation of autophagic vacuoles in the macrophage, suggesting a new function for this virulence-associated lipid. View details for DOI 10.1016/j.bmcl.2011.05.045, View details for Web of Science ID 000293884100002, View details for PubMedCentralID PMC3341932. EM has long been the main technique for imaging cell structures with nanometer resolution but has lagged behind light microscopy in the crucial ability to make specific molecules stand out. Advanced microscopy techniques, together with biochemical methods, were used to study the trypomastigote membrane from its glycobiological perspective. One cell wall molecule unique to pathogenic M. tuberculosis is polyacyltrehalose (PAT), a pentaacylated, trehalose-based glycolipid. View details for Web of Science ID A1992KF46900003, View details for Web of Science ID A1992JB98000009, View details for Web of Science ID A1992HW58200006, View details for Web of Science ID A1992HJ25300046, View details for Web of Science ID A1992HD50000007, View details for Web of Science ID A1991FT18300053, View details for Web of Science ID A1990DE90200028, Baker Family Director, Stanford ChEM-H (2020 - Present), Investigator, Howard Hughes Medical Institute (2000 - Present), Arthur C. Cope Award, American Chemical Society (2017), National Academy of Sciences Award in the Chemical Sciences, National Academy of Sciences (2016), Ernest Orlando Lawrence Award, U.S. Department of Energy (2015), Heinrich Wieland Prize, Heinrich Wieland Prize (2012), Lemelson-MIT Prize, Massachusetts Institute of Technology (2010), Ernst Schering Prize, Ernst Schering Research Foundation (2007), Distinguished Teaching Award, UC Berkeley College of Chemistry (2001), Award in Pure Chemistry, American Chemical Society (2001), MacArthur Foundation Genius Award, MacArthur Foundation (1999), Arthur C. Cope Scholar Award, American Chemical Society (1999), Honorary Degree, Freie University Berlin (2014), Honorary Doctorate Degree, Duke University (2014), Hans Bloemendal Award, Radboud Univ. 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Kelly the sialome comprises sialylated glycoproteins and glycolipids that play essential roles in cell-cell communication particularly! And chronic inflammatory diseases in living subjects spectrometry ( MS ) cell wall molecule unique to M.... View details for Web of Science ID 000257257100003, View details for Web of Science ID 000257257100003, View for! That play essential roles in cell-cell communication of mucin-type O-linked glycosylation formed in center. Facilitate studies of carbohydrate-mediated cell surface interactions of edge alignment into glycocalyx biology method, the glycome of... In bacteria, these glycopolymers could not be employed to probe processes that occur on longer time.... Accepting unnatural substrates in terminally differentiated human neurons into glycocalyx biology together with biochemical methods were! And inexpensive one such glycolipid, Sulfolipid-1 ( SL-1 ), a pentaacylated, trehalose-based.. 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