WebIn the early 1990s, Carolyn Bertozzi began mapping a glycan that attracts immune cells to lymph nodes. Sialome sweet sialome: As sialic acids are involved in many host-pathogen recognition events and are markers of embryonic and malignant tissues, there is great interest in methods for the enrichment and identification of sialylated glycoproteins from complex tissues. To enable noninvasive, real-time, spatiotemporal quantitative imaging of fatty acid fluxes in animals, we created a bioactivatable molecular imaging probe based on long-chain fatty acids conjugated to a reporter molecule (luciferin). Many developmental processes depend on proper fucosylation, but this post-translational modification is difficult to monitor in vivo. Applications to noninvasive imaging and glycoproteomic analyses are discussed. Treatment of hESCs with Ac4-5SGlcNAc during induction limited protein O-GlcNAcylation and also caused a dramatic decrease in global levels of UDP-GlcNAc. View details for Web of Science ID 000171801100023. View details for Web of Science ID 000262171100029, View details for PubMedCentralID PMC2655317. We describe a chemical strategy directed toward identifying O-GlcNAc-modified proteins from living cells or proteins modified in vitro. 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. 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. This system can be used to capture non-adherent cells specifically and to discriminate between visually similar healthy and cancerous cells in a heterogeneous ensemble based on their altered metabolic properties. [20][21], After graduating from Berkeley with a Ph.D., Bertozzi was a postdoctoral fellow at University of California, San Francisco (UCSF) with Steven Rosen, where she studied the activity of endothelial oligosaccharides in promoting cell adhesion at inflammation sites. Densely O-glycosylated mucin domains are found in a broad range of cell surface and secreted proteins, where they play key physiological roles. She was a postdoctoral fellow at the University of California, San Francisco, from 1993 to 1995. The compound was metabolized to the azidosialic acid and incorporated into cell surface and secreted sialoglycoproteins. WebCarolyn Bertozzi is the Baker Family Director of Sarafan ChEM-H, the Anne T. and Robert M. Bass Professor of Chemistry, and a professor of chemical and systems biology and of 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 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. View details for DOI 10.1074/mcp.M111.015339, View details for Web of Science ID 000306408500017. In one step, 35S-labeled by-products are then eluted from the membrane, leaving spatially separated 35S-labeled product "dots" for subsequent quantification. Formylglycine generating enzyme (FGE) recognizes a pentapeptide consensus sequence, CxPxR, and it specifically oxidizes the cysteine in this sequence to an unusual aldehyde-bearing formylglyine. Jain, M., Petzold, C. J., Schelle, M. W., Leavell, M. D., Mougous, J. D., Bertozzi, C. R., Leary, J. 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. The presence of SiaLev groups on the neuronal cell surface did not impede neurite outgrowth or significantly affect the distribution of PSA on neuronal compartments. Most cell membrane proteins are known or predicted to be glycosylated in eukaryotic organisms, where surface glycans are essential in many biological processes including cell development and differentiation. Hotsclaw, C. M., Sogi, K. M., Gilmore, S. A., Scheller, M. W., Leavell, M. D., Bertozzi, C. R., Leary, J. Laureates [3] [ edit] Laureates per country [ edit] Cell surface glycosylation is thought to be involved in barrier function against microbes at mucosal surfaces. a nanoscience research center at Lawrence Berkeley National Laboratory. Chem. The recently described O-glycoprotease OpeRATOR presents exciting opportunities for O-glycoproteomics. Subsequently, the embryos were reacted with fluorophore conjugates by means of copper-free click chemistry, enabling the visualization of glycans in vivo at subcellular resolution during development. and Shu Wang and Kim, {Hyun Jae} and Meyer, {Gerald J.} We synthesized two unnatural oligosaccharide substrates, GlcNAc beta 1-->6Gal alpha-R and Gal beta 1-->4GlcNAc beta 1-->6Gal alpha-R, that incorporate structural motifs from the native L-selection ligands into an unnatural C-glycosyl hydrocarbon scaffold. She has found a smart way to connect cells together. To elevate the reactivities of cyclooctynes, two strategies, LUMO lowering through propargylic fluorination and strain enhancement through fused aryl rings, have been explored. and Irmgard Chu Distinguished Professorship in Chemistry, UC Berkeley (2005-14), Agnes Fay Morgan Research Award, Iota Sigma Pi (2004), Fellow, American Association for the Advancement of Science (2002), Irving Sigal Young Investigator Award, Protein Society (2002), Donald Sterling Noyce Prize for Excellence in Undergraduate Teaching, UC Berkeley College of Chemistry (2001), Department of Chemistry Teaching Award, UC Berkeley (2000), Merck Academic Development Program Award, Merck (2000), Presidential Early Career Award in Science and Engineering (PECASE), The U.S. White House (2000), Camille Dreyfus Teacher-Scholar Award, Camille and Henry Dreyfus Foundation (1999), Beckman Young Investigator Award, Arnold and Mabel Beckman Foundation (1998), Glaxo Wellcome Scholar, Glaxo Wellcome (1998), Prytanean Faculty Award, Prytanean Women's Honor Society, UC Berkeley (1998), Research Innovation Award, Research Corporation (1998), Young Investigator Award, Office of Naval Research (1998), Horace S. Isbell Award in Carbohydrate Chemistry, American Chemical Society (1997), New Investigator Award in Pharmacology, Burroughs Wellcome (1997), Sloan Research Fellow, Alfred P. Sloan Foundation (1997), Pew Scholars Award in the Biomedical Sciences, Pew Charitable Trusts (1996), Young Investigator Award, Exxon Education Fund (1996), Dreyfus New Faculty Award, Camille and Henry Dreyfus Foundation (1995), Member, National Academy of Inventors (2013 - Present), Member, Institute of Medicine (2011 - Present), Member, German Academy of Sciences Leopoldina (2008 - Present), Member, National Academy of Sciences (2005 - Present), Member, American Academy of Arts and Sciences (2003 - Present), Chair, Scientific Advisory Board, Redwood Bioscience, Board Member, Board of Scientific Counslors, Broad Institutue, Board Member, Research Advisory Baord, Glaxo Smithkline, AB, Harvard University, Chemistry (1988), Membrane curvature regulates the spatial distribution of bulky glycoproteins. [reaction--see text] The synthesis of oxime-linked mucin mimics was accomplished via the incorporation of multiple ketone residues into a peptide followed by reaction with aminooxy sugars corresponding to the tumor-related T(N) and sialyl T(N) (ST(N)) antigens. Chen, Q., Zhang, D., Somorjai, G., Bertozzi, C. R. Carbohydrate sulfotransferases: mediators of extracellular communication, Chemoselective ligation reactions with proteins, oligosaccharides and cells, Inner space exploration: the chemical biologist's guide to the cell, Metabolic delivery of ketone groups to sialic acid residues - Application to cell surface glycoform engineering. We show that the enhanced analytical sensitivity enables this assay to correctly identify HIV-infected individuals otherwise missed by current OF assays. View details for Web of Science ID 000259675500001, View details for PubMedCentralID PMC2709988. Stanford chemist Carolyn Bertozzi was awarded the Nobel Prize in chemistry for her development of bioorthogonal reactions, which allow scientists to explore cells and track biological processes without disrupting the normal chemistry of the cell. This story was updated on Wednesday, Oct. 6, at 1:23 p.m. PDT. In the accompanying paper [Hemmerich, S., & Rosen, S.D. A Bioorthogonal Reaction of N-Oxide and Boron Reagents. 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. 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. The process of generating aldehyde-tagged protein followed by chemical conjugation and purification takes 20 d. View details for DOI 10.1038/nprot.2012.045, View details for Web of Science ID 000304720700005, View details for PubMedCentralID PMC3498491. We identified fluorescein-trehalose analogues that are metabolically incorporated into the trehalose mycolates of representative Mycobacterium, Corynebacterium, Nocardia, and Rhodococcus species. We find that monomers with an extended conformation first form a mobile adsorbed phase, from which they condense into amorphous clusters. This method is rapid and efficient, allowing virtually any mammalian cell to be patterned on surfaces bearing complementary DNA in under 1 h. We demonstrate this technique using several types of cells that are generally incompatible with integrin-targeting approaches, including red blood cells and primary T-cells. 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. This left Bertozzi and the rest of the lab to complete their Ph.D. work with no direct supervision. Mycobacteria are endowed with a highly impermeable mycomembrane that confers intrinsic resistance to many antibiotics. Mucin glycoproteins contribute to a wide range of cell-surface phenomena. Malignancy is associated with altered expression of glycans and glycoproteins that contribute to the cellular glycocalyx. View details for Web of Science ID 000296075300003. Recent work has implicated tyrosine sulfate as a determinant of protein-protein interactions involved in leukocyte adhesion, hemostasis and chemokine signaling. Here we show that a QC amino acid can be incorporated into a protein site-specifically using the pyrrolysine-based genetic code expansion platform, and subsequently used for ligation chemistry. These hallmarks of Mtb infection are facilitated by the metabolic adaptation of the pathogen to its surrounding environment and the biosynthesis of molecules that mediate its interactions with host immune cells. Our approach validates the use of the applied metabolic strategy to identify important functions of GalNAc-Ts in normal and pathological conditions. In this approach, metabolic labeling substrates containing bioorthogonal functional groups are provided to cells for incorporation into biopolymers by endogenous biosynthetic machinery. View details for DOI 10.1073/pnas.2335201100, View details for Web of Science ID 000187227200045, View details for PubMedCentralID PMC299823. Tests against related enzymes confirmed the compound's specificity for the UDP-GlcNAc 4-epimerase. Malaker, S. A., Shon, J., Pedram, K., Riley, N. M., Bertozzi, C. R. AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC. An RNA-centric dissection of host complexes controlling flavivirus infection. A., Cox, J. S. Noncovalent complexes of APS reductase from M-tuberculosis: Delineating a mechanistic model using ESI-FTICR MS. Gao, H., Leary, J., Carroll, K. S., Bertozzi, C. R., Chen, H. Self-assembled cellular microarrays patterned using DNA barcodes. The mineral-nucleating potential of hydroxyl groups identified here broadens the design parameters for synthetic bonelike composites and suggests a potential role for hydroxylated collagen proteins in bone mineralization. View details for Web of Science ID 000264791600009, View details for PubMedCentralID PMC2657038. The deep structural analysis enabled by this new method will enable future mechanistic studies on the biological significance of high mannose glycans on stem cell membranes and provide a general tool to examine cell surface glycosylation. 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