This template-driven mineralization technique provides an efficient approach toward bonelike composites with high mineral-hydrogel interfacial adhesion strength. The incorporation of noncanonical amino acids into recombinant proteins in Escherichia coli can be facilitated by the introduction of new aminoacyl-tRNA synthetase activity into the expression host. Acute Modulation of Mycobacterial Cell Envelope Biogenesis by Front-Line Tuberculosis Drugs. View details for DOI 10.1093/glycob/cwi064, View details for Web of Science ID 000230346400006. Senaratne, R. H., De Silva, A. D., Williams, S. J., Mougous, J. D., Reader, J. R., Zhang, T. J., Chan, S., Sidders, B., Lee, D. H., Chan, J., Bertozzi, C. R., Riley, L. W. Mucin granule intraluminal organization in living mucous/goblet cells - Roles of protein post-translational modifications and secretion. Kobertz, W. R., Bertozzi, C. R., Bednarski, M. D. Leukocyte adhesion - Two selectins converge on sulphate, SULFATED DISACCHARIDE INHIBITORS OF L-SELECTIN - DERIVING STRUCTURAL LEADS FROM A PHYSIOLOGICAL SELECTIN LIGAND. A computational model predicted that these glycoproteins would influence transmembrane receptor spatial organization and function. 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. A predominantly hydrophobic aglycone site facilitates accommodation of a variety of 2-linked sialosides; a versatility that offers the potential for glycan hydrolysis across a range of biological and technological platforms. Isotopic recoding is achieved via metabolic incorporation of a defined mixture of N-acetylglucosamine isotopologs into N-glycans. Finally, we engineered the Golgi-resident glycosyltransferase FucT7 for tunable control by Tmp-SLF in mammalian cells. Bioorthogonal tools enable cell-type-specific proteomics, a prerequisite to understanding biological processes in multicellular organisms. Human embryonic stem cells were found to have high levels of high mannose glycans, which contrasts with IMR-90 fibroblasts and a human normal breast cell line, where complex glycans are by far the most abundant and high mannose glycans are minor components. A cell metabolic labeling experiment can be completed in approximately 4 d. View details for DOI 10.1038/nprot.2007.422, View details for Web of Science ID 000253140000033. She is also an Investigator at the Howard Hughes Medical Institute (HHMI) and is the former Director of the Molecular Foundry. The controlled integration of organic and inorganic components confers natural bone with superior mechanical properties. These results show that CysH is important for Mtb protection during the chronic infection phase, and that resistance to nitrosative and oxidative stress may be the mechanism of this protection. The vast majority of H. ducreyi strains contain high levels of sialic acid (N-acetylneuraminic acid, NeuAc) in their LOS. [3] At Stanford University, she holds the Anne T. and Robert M. Bass Professorship in the School of Humanities and Sciences. Overall, our findings provide a quantitative characterization of O-GlcNAc glycoproteins and their corresponding modification sites in primary human T cells, which will facilitate mechanistic studies into the function of O-GlcNAc in T cell activation. Poly-alpha2,8-sialic acid (PSA) has been implicated in numerous normal and pathological processes, including development, neuronal plasticity, and tumor metastasis. Modification of the terminal arabinan residues of this lipoglycan with mannose caps in M. tuberculosis or with phosphoinositol caps in Mycobacterium smegmatis results in distinct host immune responses. [47] Lycia Therapeutics focuses on developing technology which utilizes lysosome-targeting chimeras (LYTACs). With this technique, we studied the dynamics of glycan trafficking and identified a population of sialoglycoconjugates with unexpectedly rapid internalization kinetics. The assay was validated by measuring K(M) values for PAPS and K(I) values for PAP, the product of sulfuryl transfer. Here, we describe the broad application of the method to analyze glycoproteomes from a collection of tissue-diverse cell lines. In addition, similar sulfated epitopes are known to be expressed on HEV-like vessels of chronically inflamed tissues; indicating that this sulfotransferase may also contribute to inflammatory lymphocyte recruitment. View details for DOI 10.1016/S0166-6851(03)00196-8, View details for Web of Science ID 000185793400002. In conclusion, the new ADAP assay can reliably detect the three cardinal islet autoantibodies/antibodies in 1muL serum with high sensitivity. The Symbol Nomenclature for Glycans (SNFG) is a community-curated standard for the depiction of monosaccharides and complex glycans using various colored-coded, geometric shapes, along with defined text additions. Several anti-tuberculosis drugs target the biogenesis of this complex envelope, but their efficacy is declining due to resistance. Herein we analyzed the glycopeptide substrate preferences of several ppGalNAcT family members using a library screening approach. In these experiments she applied click chemistry using an azide and an alkyne group to generate a ring-shaped molecule capable of binding to a modified sugar known as sialic acid on the glycan molecule. Bertozzi has applied the techniques of bioorthogonal chemistry to study glycocalyx, the sugars that surround the cell membrane. Rv3406 was identified based on its homology to the alkyl sulfatase AtsK from Pseudomonas putida. Fluorescent tagging in cultured cells and developing organisms has revealed important insights into the dynamics of these structures during growth and development. In eukaryotic sulfatases, an active site cysteine residue is oxidized to the aldehyde-containing C(alpha)-formylglycine residue by the formylglycine-generating enzyme (FGE). View details for DOI 10.1007/s00216-016-9934-9, View details for Web of Science ID 000391364200021. We describe here a new ELISA that measures the binding between selectin-IgG chimeras and a physiological ligand for L-selectin and can be used to screen selectin inhibitors. In this paper, an efficient enzyme kinetics assay for Stf0 using electrospray ionization (ESI) mass spectrometry is presented. View details for Web of Science ID 000207584000003, View details for PubMedCentralID PMC138933. View details for DOI 10.1073/pnas.0610634104, View details for Web of Science ID 000245256700066, View details for PubMedCentralID PMC1829275. Inducible genetic inhibition of angiogenesis and Vegf signaling during granuloma formation results in bacterial growth deficits. ADAP detects zepto- to attomoles of antibodies in 2 L of sample with a dynamic range spanning 5-6 orders of magnitude. View details for Web of Science ID 000243895200017. The omega-alkynyl-myristate and -palmitate analogs used with click chemistry and azido-probes will be invaluable to study protein acylation in vitro, in cells, and in vivo. Radioimmunoassay remains the gold standard for these challenging antibody biomarkers, but the limited multiplexability and reliance on hazardous radioactive reagents have prevented their use outside specialized testing facilities. Analysis of the growth kinetics leads to a quantitative model in which tetramer creation is rate limiting. Azides installed within cell surface glycoconjugates by metabolism of a synthetic azidosugar were reacted with a biotinylated triarylphosphine to produce stable cell-surface adducts. This bacterial enzyme purified from Akkermansia muciniphila cleaves N-terminally to serine and threonine residues that are modified with (preferably asialylated) O-glycans. View details for Web of Science ID 000168932900013. Gerling-Driessen, U. M., Tomlin, F. M., Cambier, C. J., Liu, Y., Bertozzi, C. R. A Model for Accelerating Patient-to-Bench Research. A., Bertozzi, C. R. Compositional profiling of heparin/heparan sulfate using mass spectrometry: assay for specificity of a novel extracellular human endosulfatase. WebBio BIO Professor Carolyn Bertozzi's research interests span the disciplines of chemistry and biology with an emphasis on studies of cell surface sugars important to human health and disease. Upon exposure to mycobacterial cell wall lipids, 166 macrophage proteins showed differential expression. Changes in glycosylation are often a hallmark of disease states. [36] This Pharmaceuticals dissolved in 2005. The ability to chemoselectively monitor H(2)O(2) fluxes in real time in living animals offers opportunities to dissect H(2)O(2)'s disparate contributions to health, aging, and disease. Using this approach quantities of homogeneous material were obtained for structural and functional analysis. Protein glycosylation is a heterogeneous post-translational modification (PTM) that plays an essential role in biological regulation. We determined the crystal structure of the apo Rv3406 sulfatase at 2.5 . We frame central issues regarding determination of protein-level variation and PTMs, including some paradoxes present in the field today. The selectins are a family of three adhesion molecules (L-, P-, and E-) that direct the interaction of circulating leukocytes with endothelial cells during the first step in recruitment to tissue sites. This review highlights the development of bioorthogonal chemical reporters and reactions and their application in living systems. View details for DOI 10.1371/journal.pone.0242049. Sialylation, sulfation, and fucosylation appear to be required for the avid interaction of this ligand with L-selectin, but the exact carbohydrate structures involved in recognition remain undefined. Application of our enzymatic toolkit to ascites fluid and tissue slices from patients with ovarian cancer facilitated characterization of patients based on differences in mucin cleavage and expression patterns. Both reactive partners are abiotic and chemically orthogonal to native cellular components. Electron-based dissociation methods are necessary to capture the O-glycopeptide diversity present in OpeRATOR digestions. One of the endothelial-derived ligands for L-selectin is GlyCAM-1 (previously known as Sgp50), a mucin-like glycoprotein with sulfated, sialylated, and fucosylated O-linked oligosaccharide chains. [reaction: see text] Here we report a novel modification of our previously reported "Staudinger ligation" that generates an amide bond from an azide and a specifically functionalized phosphine. These metabolites are products of the sulfate assimilation pathway. [38] Redwood Bioscience is a biotechnology company that uses SMARTag, a site-specific protein modification technology that allows small drugs to attach to sites on the proteins and can be used to help fight cancers. Using an Orbitrap Fusion Tribrid mass spectrometer, we identified 699 intact glycopeptides from 192 glycoproteins. Several protein lysine methyltransferases (PKMTs) modify histones to regulate chromatin-dependent cellular processes, such as transcription, DNA replication and DNA damage repair. View details for DOI 10.1002/anie.200806319, View details for Web of Science ID 000266415400022, View details for PubMedCentralID PMC2868584. 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. Physical Principles of Membrane Shape Regulation by the Glycocalyx. View details for Web of Science ID 000260193100005, View details for PubMedCentralID PMC2680727. View details for DOI 10.1021/jacs.8b03074, View details for Web of Science ID 000435525500001, View details for DOI 10.3389/fmicb.2018.01117, View details for Web of Science ID 000433326300001. Phagocytosis is the central process by which macrophage cells internalize and eliminate infectious microbes as well as apoptotic cells. Lu, C., Jones, T., Pedram, K., Bertozzi, C., Paszek, M., Cui, B. We report the cloning, expression, and characterization of the first UDP-GalNAc:polypetide N-acetylgalactosaminyltransferase (ppGalNAc-T) from the human disease-causing parasite, Toxoplasma gondii. Conversely, unsialylated LacNAc glycans, enriched in the epithelial ducts, sequestered Gal-1 in the extracellular environment, ultimately attenuating invasive potential. 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. Shieh, P., Siegrist, M. S., Cullen, A. J., Bertozzi, C. R. Glycocalyx engineering reveals a Siglec-based mechanism for NK cell immunoevasion. She is an elected member of the National Academy of Sciences, the American Academy of Arts and Sciences, and the German Academy of Sciences Leopoldina. Sensitive detection of multiple islet autoantibodies in type 1 diabetes using small sample volumes by agglutination-PCR. Alterations in GalNAc-Ts expression were shown to provide huge variability in the O-glycoproteome in various pathologies, including cancer. Li phin vn-chng si to pan-lan-chhan , ng cho-tet yung phin-si fet-ch khok-chhng kh ke nui-yng. Carolyn Bertozzi, PhD, was one of three scientists to be awarded the 2022 Nobel Prize in Chemistry earlier today for the development of click chemistry and bio-orthogonal chemistry. Eliminate infectious microbes as well as apoptotic cells, enriched in the epithelial,! 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