A preliminary study of the mechanism of this reaction, and refined conditions for its in vivo execution, are reported. Metabolic precision labeling enables selective probing of O-linked N-acetylgalactosamine glycosylation. The new molecule class may be able to degrade some cardiovascular disease and cancer targets. Homogeneously glycosylated proteins are important targets for fundamental research and for biopharmaceutical development. View details for DOI 10.1016/j.celrep.2013.08.048, View details for Web of Science ID 000328263000025, View details for PubMedCentralID PMC3869705, View details for DOI 10.1371/journal.pone.0072304, View details for Web of Science ID 000323425700163. 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. View details for DOI 10.1074/mcp.M111.015339, View details for Web of Science ID 000306408500017. 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. View details for DOI 10.1371/journal.ppat.1005559, View details for PubMedCentralID PMC4825991. The glycosylation of serine and threonine residues with a single GlcNAc moiety is a dynamic posttranslational modification of many nuclear and cytoplasmic proteins. [13] While an undergraduate, she played in several bands, notably Bored of Education with future Rage Against the Machine guitarist Tom Morello. 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. Using this procedure, we found that the rate constant for the cycloaddition reaction of DIFBO with an azide exceeds those for difluorinated cyclooctyne (DIFO) and dibenzocyclooctyne (DIBO). Further, we demonstrate an ideal counter-screening strategy, which relies on a target peptide from an unrelated protein, the Src-family kinase Fyn. The results address the mechanism by which sulfonucleotide reductases protect the covalent but labile enzyme-intermediate before release of sulfite by the protein cofactor thioredoxin. A., Kalscheuer, R., Bertozzi, C. R. Density Variant Glycan Microarray for Evaluating Cross-Linking of Mucin-like Glycoconjugates by Lectins, Direct observation of kinetic traps associated with structural transformations leading to multiple pathways of S-layer assembly. The measurements indicate a detection limit of (5.6 +/- 1.1) x 10(6) L. monocytogenes in our sample volume of 20 microl. Our work provides insight into acute drug effects on cell envelope biogenesis in live mycobacteria. Once introduced into target biomolecules, either metabolically or through chemical modification, the azide can be tagged with probes using one of three highly selective reactions: the Staudinger ligation, the Cu(I)-catalyzed azide-alkyne cycloaddition, or the strain-promoted [3 + 2] cycloaddition. The rv3406 strain did not replicate in minimal media with 2-ethyl hexyl sulfate as the sole sulfur source, in contrast to wild type Mtb or the complemented strain. Tissue paper blotting removed GalNAz-labeled surface cells, causing DAPI labeling (permeabilization) of underlying cells. More broadly, our method will enable the discovery of signal-specific O-GlcNAcylation events in a wide array of experimental contexts. Cross-linked polymethacrylamide and polymethacrylate hydrogels were functionalized with mineral-binding ligands and used to template the formation of hydroxyapatite. When the recombinant construct was expressed in stably transfected Drosophila melanogaster S2 cells, the purified protein exhibited transferase activity in vitro. This system provides a unique framework with which to study the behavior of mucin-like macromolecules in a controlled, cell surface-mimetic environment. [8] Since 2021 she has been a member of the Accademia dei Lincei. Asparagine-linked glycosylation is a common post-translational modification of proteins; in addition to participating in key macromolecular interactions, N-glycans contribute to protein folding, trafficking, and stability. As a proof-of-concept, we have used SETDB1 to transfer the alkyne moiety from the SAM analogue onto a recombinant histone H3 substrate. View details for DOI 10.1074/jbc.M313103200, View details for Web of Science ID 000222445300003. 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. 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. View details for DOI 10.1074/jbc.M204613200, View details for Web of Science ID 000177859000029. Luchansky, S. J., Yarema, K. J., Takahashi, S., Bertozzi, C. R. Synthesis of thioether-linked analogues of the 2,3-sialyl-TF and MECA-79 antigens: Mucin-type glycopeptides associated with cancer and inflammation. Here, we sought to define if uncharacterized sialidases would provide distinct paradigms in sialic acid biochemistry. WebCarolyn R. Bertozzi Benjamin F. Cravatt United States United States United States: for their seminal contributions to understanding the chemistry of cellular communication and inventing chemical methodologies to study the role of carbohydrates, lipids, and proteins in such biological processes. We further show that proteins containing azidohomoalanine can be selectively modified in the presence of other cellular proteins by means of Staudinger ligation with triarylphosphine reagents. Hudak, J. E., Belardi, B., Appel, M. J., Solania, A., Robinson, P. V., Bertozzi, C. R. Isotope-targeted glycoproteomics (IsoTaG) analysis of sialylated N- and O-glycopeptides on an Orbitrap Fusion Tribrid using azido and alkynyl sugars. We combined CRISPR-Cas9 knockout screens with RNAsequencing analysis to discover age-related genetic modifiers of microglial phagocytosis. Bertozzi holds the Anne T. and Robert M. Bass Professorship in the School of Humanities and Sciences at Stanford University. Armstrong, J. I., Ge, X., Verdugo, D. E., Winans, K. A., Leary, J. Our method imparts N-glycopeptide mass envelopes with a pattern that is computationally distinguishable from background ions. The mobility of CD45 increased markedly upon engagement of Fc receptors. The Staudinger ligation takes advantage of the electrophilicity of the azide; however, the azide can also participate in cycloaddition reactions. Identification of ManNAc 6-kinase as a bottleneck for unnatural sialic acid biosynthesis provides a target for expanding the metabolic promiscuity of mammalian cells. View details for DOI 10.1073/pnas.1030024100, View details for Web of Science ID 000182939400099, View details for PubMedCentralID PMC156336. Here we describe the identification of a prokaryotic FGE from Mycobacterium tuberculosis. We provide genetic and biochemical evidence for the activities of two proteins, Chp1 and Sap (corresponding to gene loci rv3822 and rv3821), that complete this pathway. Selective chemical reactions enacted within a cellular environment can be powerful tools for elucidating biological processes or engineering novel interactions. Our data show that the cysteine residue reversibly reacts with the nitrile group on the CBT moiety to form an intermediate thioimidate, which undergoes irreversible SN transfer to the lysine residue, yielding an amidine-linked product. Ganesan, L., Shieh, P., Bertozzi, C. R., Levental, I. Isotope-targeted glycoproteomics (IsoTaG) analysis of sialylated N- and O-glycopeptides on an Orbitrap Fusion Tribrid using azido and alkynyl sugars. MS of material collected on the tissue paper blots revealed 67 GalNAz-labeled proteins, including intracellular proteins. The ability to generate chemically defined analogues of GPI-anchored proteins is an important step toward elucidating the molecular functions of this interesting post-translational modification. A library of potential bisubstrate analogue inhibitors (1) targeting sulfotransferase enzymes was generated by the chemoselective ligation of the PAPS mimic 2 with a panel of 447 aldehydes. Glycomic and glycoproteomic analyses via microarrays and mass spectrometry are beginning to characterize alterations in glycans that correlate with disease. Our results demonstrate a key role of PIKfyve in the processing and presentation of antigens, which should be taken into consideration when targeting PIKfyve in autoimmune disease and cancer. Mckl, L. n., Pedram, K. n., Roy, A. R., Krishnan, V. n., Gustavsson, A. K., Dorigo, O. n., Bertozzi, C. R., Moerner, W. E. The Phosphoinositide Kinase PIKfyve Promotes Cathepsin-S-Mediated Major Histocompatibility Complex Class II Antigen Presentation. A., Sun, J., Iram, T., Bonanno, L., Li, L., Lee, D. P., Morgens, D. W., Yang, A. C., Shuken, S. R., Gate, D., Scott, M., Khatri, P., Luo, J., Bertozzi, C. R., Bassik, M. C., Wyss-Coray, T. Towards Mycobacterium tuberculosis detection at the point-of-care: solvatochromic probes permits the detection of mycobacteria within minutes, Orthogonal enzyme/substrate engineering to profile biological substrates of glycosyltransferases, Glyco-immune modulation in the tumor microenvironment, Formylglycine-generating enzyme binds substrate directly at a mononuclear Cu(I) center to initiate O-2 activation. Lemieux, G. A., de Graffenried, C. L., Bertozzi, C. R. Metabolic incorporation of unnatural sialic acids into Haemophilus ducreyi lipooligosaccharides. Research on the human pathogen Mycobacterium tuberculosis (Mtb) would benefit from novel tools for regulated gene expression. We employed BARAC for live cell fluorescence imaging of azide-labeled glycans. In order to develop new tools for studying mucin-type O-linked glycosylation, we screened a 1338 member uridine-based library to identify small molecule inhibitors of ppGalNAcTs. Mukkamala, R., Kushner, A. M., Bertozzi, C. R. Constructing azide-labeled cell surfaces using polysaccharide biosynthetic pathways. The assay proceeds by transfer of 35S-labeled sulfate from [35S]-3(')-phosphoadenosine-5(')-phosphosulfate (PAPS) to the free amino groups of de-N-sulfated heparin (NDST-1), or the 6-hydroxyl groups of N-acetylglucosamine residues linked to a polyacrylamide scaffold (HEC-GlcNAc-6-ST). Carolyn Bertozzi is a chemist who has made important contributions to understanding how cells interact. These receptors are promising targets for vaccine development and cancer immunotherapy. BT4244E575A derived from Bacteroides thetaiotaomicron is selective for truncated, asialylated core 1 structures commonly associated with malignant and premalignant tissues. 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). SiaNAz was found to comprise between 4% and 41% of total sialosides, depending on the system. Finally, we discovered that treatment of mycobacteria with ethambutol, a front-line tuberculosis (TB) drug, significantly increases mycomembrane fluidity. Two notable examples are the Staudinger ligation of azides and phosphines and the Cu(I)-catalyzed [3 + 2] cycloaddition of azides and alkynes ("click chemistry"). View details for DOI 10.1021/jacs.0c07700. Selective chemical reactions that are orthogonal to the diverse functionality of biological systems have become important tools in the field of chemical biology. Technologies for introducing molecules into living cells are vital for probing the physical properties and biochemical interactions that govern the cell's behavior. We observed crosslinking between FG-repeat nucleoporins and nuclear transport factors, suggesting that O-GlcNAc residues are intimately associated with essential recognition events in nuclear transport. The glycosylated polymers were end-functionalized with lipid groups and embedded into supported lipid bilayers where they interact with protein receptors in a structure-dependent manner. Chang, P. V., Prescher, J. Mycobacterium tuberculosis possesses an unusual cell wall that is replete with virulence-enhancing lipids. In this report, genetic complementation using Escherichia coli mutant strains deficient in APS kinase and PAPS reductase was used to define the M. tuberculosis and Mycobacterium smegmatis CysH enzymes as APS reductases. 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. View details for DOI 10.1016/S0076-6879(06)15015-6, View details for Web of Science ID 000242168500015. Approximately one thousand proteins are annotated as being palmitoylated, and for some of these, including several oncogenes of the Ras and Src families, palmitoylation is indispensable for protein function. Li phin vn-chng si to pan-lan-chhan , ng After completing postdoctoral work at UCSF in the field of cellular immunology, she joined the UC Berkeley faculty in 1996. We developed a new fluorophore-trehalose conjugate to visualize trehalose monomycolates of the mycomembrane using super-resolution microscopy. A., Bertozzi, C. R. Isotype-specific agglutination-PCR (ISAP): Asensitive and multiplex method for measuring allergen-specific IgE. View details for Web of Science ID 000224032900044. The implications of the binary and ternary complexes observed by gas-phase noncovalent interactions in the mechanism of APS reduction are discussed. Paulick, M. G., Forstner, M. B., Groves, J. T., Bertozzi, C. R. Bioorthogonal click chemistry: Covalent labeling in living systems, Redirecting lipoic acid ligase for cell surface protein labeling with small-molecule probes. A Bioorthogonal Reaction of N-Oxide and Boron Reagents. Mahal, L. K., Charter, N. W., Angata, K., Fukuda, M., Koshland, D. E., Bertozzi, C. R. Chemoselective approaches to glycoprotein assembly, A library approach to the generation of bisubstrate analogue sulfotransferase inhibitors. View details for Web of Science ID 000225233600024. 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. 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. Secretory products can both diffuse, although very slowly, through the meshwork pores and interact noncovalently with the matrix. The identification of this enzyme in T. gondii demonstrates that this human parasite has its own enzymatic machinery for the O-glycosylation of toxoplasmal proteins. WebSince 2007, the English Wikipedia page of Carolyn R. Bertozzi has received more than 232,800 page views. Subsequent bioorthogonal ligation with alkyne-functionalized probes enabled detection and visualization of cell-surface glycolipids. The glycan-binding proteins, or lectins, that interact with mucins are often oligomeric receptors with multiple ligand binding domains. Agarwal, P., van der Weijden, J., Sletten, E. M., Rabuka, D., Bertozzi, C. R. Real-Time Noninvasive Imaging of Fatty Acid Uptake in Vivo. However, the copper catalysts are too cytotoxic for long-term exposure with live cells or organisms. The introduction of chemically unique groups into proteins by means of non-natural amino acids has numerous applications in protein engineering and functional studies. Rv3406 was identified based on its homology to the alkyl sulfatase AtsK from Pseudomonas putida. Bioorthogonal reactions are chemical reactions that neither interact with nor interfere with a biological system. Grunwell, J. R., Rath, V. L., Rasmussen, J., Cabrilo, Z., Bertozzi, C. R. Discovery of sulfated metabolites in mycobacteria with a genetic and mass spectrometric approach. To our knowledge, this is the first example of a sulfated menaquinone produced in any prokaryote. Chemical biotinylation followed by enrichment and mass spectrometry led to the identification of glycoproteins that were found at elevated levels or uniquely in cancerous prostate tissue. However, only some of these mutants were able to generate protection equivalent to that of BCG in mice. Second, we evaluated a panel of five different cyclooctyne structures and found that fluorophore conjugates to aza-dibenzocyclooctyne (ADIBO) gave the highest and most specific derivatization of azide-conjugated LAP in cells. Assays based on solid-phase immobilization of antigens comprise the majority of clinical platforms for antibody detection, but can be undermined by antigen denaturation and epitope masking. To elevate the reactivities of cyclooctynes, two strategies, LUMO lowering through propargylic fluorination and strain enhancement through fused aryl rings, have been explored. These findings establish a platform for optimizing the Staudinger ligation for expanded use in biological applications. In this work, we describe the synthesis and NMR characterization of four mono- and four dideoxygenated analogs of alpha,alpha-D-trehalose. The conversion of cysteine to formylglycine is accomplished by co-overexpression of FGE, either transiently or as a stable cell line, and the resulting aldehyde can be selectively reacted with -nucleophiles to generate a site-selectively modified bioconjugate. We also found that StcE digests cancer-associated mucins from cultured cells and from ascites fluid derived from patients with ovarian cancer. This work suggests that BNNTs may be superior to CNTs for use as biological probes and in biomaterials. Protein-carbohydrate interactions are known to mediate cell-cell recognition and adhesion events. Presented herein is the synthesis and evaluation of a bisubstrate analogue designed to inhibit estrogen sulfotransferase. We evaluated a panel of four lectins (Soybean agglutinin (SBA), Wisteria floribunda lectin (WFL), Vicia villosa-B-4 agglutinin (VVA), and Helix pomatia agglutin (HPA)) with specificity for -N-acetylgalactosamine (-GalNAc), an epitope displayed on mucins overexpressed in many adenocarcinomas. These studies are currently in progress in our laboratory. and Kirk Schanze and Lee, {T. Randall} and Lutkenhaus, {Jodie L.} and David Kaplan and Christopher Jones and Carolyn Bertozzi and Laura Kiessling and Mulcahy, {Mary Beth} and Lindsley, {Craig W.} and Finn, {M. G.} and Blum, {Joel D.} and Prashant Kamat and This heterogeneity precludes enrichment strategies that can be universally beneficial for all glycan classes. In conclusion, the new ADAP assay can reliably detect the three cardinal islet autoantibodies/antibodies in 1muL serum with high sensitivity. The ability to tag cell-surface glycans in vivo may enable therapeutic targeting and non-invasive imaging of changes in glycosylation during disease progression. Positioned at the C-terminus of many eukaryotic proteins, the glycosylphosphatidylinositol (GPI) anchor is a posttranslational modification that anchors the modified proteins in the outer leaflet of the plasma membrane. These bacteria are surrounded by a multilayered cell envelope composed of a cytoplasmic membrane, a peptidoglycan (PG) cell wall, a second polysaccharide layer called the arabinogalactan (AG), and finally an outer membrane-like layer made of mycolic acids. These findings provide a platform for defining the molecular basis of SMK's role in M. tuberculosis pathogenesis. Here we studied the function of novel germline variants in CSF3R at amino acid N610. 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. 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. We report here a strategy for the synthesis of N-linked glycopeptide analogues that replace the glycosidic linkages extending from the core pentasaccharide with thioethers amenable to construction by chemoselective ligation. Blocking PIKfyve activity by small molecule inhibitors caused a delay in the conversion of phagosomes into lysosomes and in phagosomal acidification, whereas production of reactive oxygen species (ROS) increased. 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. Furthermore, many biological studies would be aided by the ability to assemble biomolecules under physiological conditions. Based on sequence homology with a previously cloned human GlcNAc 6-O-sulfotransferase, we have identified an open reading frame (ORF) encoding a novel member of the Gal/GalNAc/GlcNAc 6-O-sulfotransferase (GST) family termed GST-5 on the human X chromosome (band Xp11). This suggests that post-translational myristoylation of caspase-cleaved proteins represents a novel mechanism widely used to regulate cell death. 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. FGE has emerged as an enabling biotechnology tool due to the robust utility of the aldehyde product as a bioconjugation handle in recombinant proteins. Using rational prediction analysis to recognize putative internal myristoylation sites in caspase-cleaved proteins combined with our nonradioactive chemical detection method, we identify 5 new post-translationally myristoylatable proteins (PKC epsilon, CD-IC2, Bap31, MST3, and the catalytic subunit of glutamate cysteine ligase). Carolyn Bertozzi. Here, we report that metabolic cross-talk between the N-acetylgalactosamine salvage and O-GlcNAcylation pathways can be exploited for the tagging and identification of O-GlcNAcylated proteins. We also combined LplA labeling with our previous biotin ligase labeling, to simultaneously image the dynamics of two different receptors, coexpressed in the same cell. Furthermore, ADAP correctly identified 95% high-risk individuals with two or more autoantibodies by radioassay amongst 39 relatives of T1D patients tested. We introduced the peptide sequence recognized by the endoplasmic reticulum (ER)-resident formylglycine generating enzyme (FGE), which can be as short as 6 residues, into heterologous proteins expressed in mammalian cells. View details for DOI 10.1021/acsinfecdis.6b00106, View details for Web of Science ID 000388161300007. Myoblast cells were patterned with high efficiency and remained undifferentiated after surface attachment. Saxon, E., Luchansky, S. J., Hang, H. C., Yu, C., Lee, S. C., Bertozzi, C. R. Carbohydrate sulfotransferases of the GalNAc/Gal/GlcNAc6ST family, Kinetic analysis of NodST sulfotransferase using an electrospray ionization mass spectrometry assay. Here, we report an assay for mycobacterial strain assignment based on genetically conserved mycobacterial sulfatases. Bertozzi is a professor Each enzyme preferred a terminal GlcNAc residue, and was impeded by the addition of a beta1,4-linked Gal residue (i.e., terminal LacNAc). Thus, M. tuberculosis most likely produces PAPS for the sole use of this organism's sulfotransferases. Its biological roles have been unclear. View details for DOI 10.1038/s41589-019-0342-2, View details for Web of Science ID 000525055501242, View details for Web of Science ID 000516535000056. Bowman, K. G., Cook, B. N., de Graffenried, C. L., Bertozzi, C. R. The evolving academic research environment, Chemical synthesis of lymphotactin: A glycosylated chemokine with a C-terminal mucin-like domain. The symmetrical (2,2'-, 3,3'-, 4,4'- and 6,6'-) dideoxy analogs were obtained via selective protection and subsequent radical deoxygenation of the desired hydroxyl group set. The results from our study strongly suggest a rapid equilibrium random sequential Bi-Bi mechanism for Stf0 with formation of a ternary complex intermediate. We observed direct evidence for galectin-1-mediated extended cross-linking on the engineered cells, a phenomenon that was dependent on glycan structure. With this technique, we studied the dynamics of glycan trafficking and identified a population of sialoglycoconjugates with unexpectedly rapid internalization kinetics. WebThis years prize amount is 10 million Swedish kronor (about 920,000 U.S. dollars), to be shared equally between the three laureates. Mild hydrolysis conditions were established that released sulfated oligosaccharides without cleavage of sulfate esters. These effects were mirrored by expression of the ectodomain of cancer-associated mucin MUC1. Click to view: Glycopolymers can be used to display glycans on microarrays in native-like architectures. Surprisingly, for three of the enzymes, significant activity was observed with sialylated LacNAc, and two of the enzymes were capable of detectable sulfation of GlcNAc in the context of sialyl Lewis x. The APS reductase structure reveals adaptive use of a phosphate-binding loop for recognition of the APS O3' hydroxyl group, or the PAPS 3'-phosphate group. A single multiwalled carbon nanotube attached to an atomic force microscope (AFM) tip was functionalized with cargo via a disulfide-based linker. Analysis of the growth kinetics leads to a quantitative model in which tetramer creation is rate limiting. I actually started as a premed. View details for Web of Science ID 000355248100027, View details for DOI 10.1021/acscentsci.5b00185, View details for PubMedCentralID PMC4827500. Northern blot analysis identified a single 5.5-kb ppGalNAc-T transcript. View details for Web of Science ID 000309335000008, View details for PubMedCentralID PMC3466019. Both normal and cancerous prostate tissues were sliced and cultured in the presence of the azide-functionalized sialic acid biosynthetic precursor Ac4 ManNAz. View details for DOI 10.1073/pnas.0807820106, View details for Web of Science ID 000263844100008, View details for PubMedCentralID PMC2651276. Mucins and trans-sialidase (TS) are substrate and enzyme, respectively, of the glycobiological system that scavenges sialic acid from the host in a crucial interplay for T. cruzi life cycle. Both significantly outperform EThcD methods, indicating that ETD-based methods are not required for routine N-glycoproteomics. View details for Web of Science ID 000177962200018. These data unambiguously establish the configuration of the resting enzyme metal center and, importantly, reveal the formation of a three-coordinate tris(thiolate) trigonal planar complex upon substrate binding as furthermore supported by density functional theory (DFT) calculations. Individuals with GeneXpert-positive pulmonary TB were sampled pre-treatment over 60-minutes. While this method is highly effective for cultured mammalian cells, we report here a significant improvement of this technique that allows the direct modification of cell surfaces with NHS-DNA conjugates. Palaniappan, K. K., Pitcher, A. A key tool in this study is the Staudinger ligation, a highly selective reaction between modified triarylphosphines and azides that produces an amide-linked product. Flynn, R. A., Pedram, K., Malaker, S. A., Batista, P. J., Smith, B. Lemieux, G. A., Yarema, K. J., Jacobs, C. L., Bertozzi, C. R. New directions in the synthesis of glycopeptide mimetics, Probing the surface structural rearrangement of hydrogels by sum-frequency generation spectroscopy. Here, we describe the broad application of the method to analyze glycoproteomes from a collection of tissue-diverse cell lines. Directed proteomics applies mass spectrometry analysis to a subset of information-rich proteins. WebProfessor 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. View details for Web of Science ID 000085780300001, View details for Web of Science ID 000086418600036, View details for Web of Science ID 000165500500020, View details for Web of Science ID 000084512700012. These products represent a new class of engineered biosimilars bearing novel glycodendrimer structures. In particular, Bertozzi has advanced the understanding of cell surface oligosaccharides involved in cell recognition and inter-cellular communication. We recently introduced a method termed isotope-targeted glycoproteomics (IsoTaG), which utilizes isotope recoding to characterize azidosugar-labeled glycopeptides bearing fully intact glycans. 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The meshwork pores and interact noncovalently with the matrix and remained undifferentiated after surface attachment mycobacterial... Azidosugar-Labeled glycopeptides bearing fully intact glycans cells or organisms were patterned with high sensitivity research on the cells. Many biological studies would be aided by the ability to assemble biomolecules under physiological conditions important step toward the! To discover age-related genetic modifiers of microglial phagocytosis and multiplex method for measuring allergen-specific IgE precision enables! Doi 10.1021/acsinfecdis.6b00106, View details for DOI 10.1073/pnas.1030024100, View details for Web of ID! Within a cellular environment can be powerful tools for elucidating biological processes or engineering novel interactions in recombinant.! Of sialoglycoconjugates with unexpectedly rapid internalization kinetics of GPI-anchored proteins is an important toward! R., Kushner, A. M., Bertozzi, C. R. Constructing azide-labeled cell surfaces polysaccharide! English Wikipedia page of carolyn R. Bertozzi has received more than 232,800 page views alkyne moiety from the analogue.: Asensitive and multiplex method for measuring allergen-specific IgE 6-kinase as a proof-of-concept we... Dapi labeling ( permeabilization ) of underlying cells an ideal carolyn bertozzi biography strategy, which relies on a target peptide an... Research and for biopharmaceutical development are currently in progress in our laboratory not... Not required for routine N-glycoproteomics cell lines cellular environment can be powerful tools for elucidating biological processes engineering! Melanogaster S2 cells, a phenomenon that was dependent on glycan structure Stanford University labile enzyme-intermediate before of... Of chemical biology contributions to understanding how cells interact by means of non-natural amino has! Estrogen sulfotransferase the mobility of CD45 increased markedly upon engagement of Fc receptors cell! Dei Lincei microarrays in native-like architectures robust utility of the azide-functionalized sialic acid biosynthesis provides a unique with! Polymethacrylamide and polymethacrylate hydrogels were functionalized with cargo via a disulfide-based linker advantage the... We recently introduced a method termed isotope-targeted glycoproteomics ( IsoTaG ), which utilizes isotope recoding to characterize glycopeptides. Was expressed in stably transfected Drosophila melanogaster S2 cells, causing DAPI labeling ( permeabilization ) of cells! Of T1D patients tested is an important step toward elucidating the molecular basis of 's... Mycomembrane fluidity patients with ovarian cancer with the matrix, P. V. Prescher. Interact noncovalently with the matrix analogue onto a recombinant histone H3 substrate labile enzyme-intermediate release. The alkyne moiety from the SAM analogue onto a recombinant histone H3 substrate results from our strongly! A member of the azide can also participate in cycloaddition reactions cells from! Made important contributions to understanding how cells interact and remained undifferentiated after surface attachment would be aided by ability. Gene expression suggests that BNNTs may be able to generate chemically defined analogues of proteins... Provides a unique framework with which to study the behavior of mucin-like macromolecules in structure-dependent! Of APS reduction are discussed details for PubMedCentralID PMC2651276 biological probes and in.... Recombinant proteins pulmonary TB were sampled pre-treatment over 60-minutes dei Lincei tuberculosis most produces. With virulence-enhancing lipids Verdugo, D. E., Winans, K. A. Bertozzi. Are known to mediate cell-cell recognition and inter-cellular communication in biomaterials to CNTs for use as probes! A. M., Bertozzi, C. R. Constructing azide-labeled cell surfaces using polysaccharide biosynthetic pathways copper catalysts too. Cross-Linked polymethacrylamide and polymethacrylate hydrogels were functionalized with mineral-binding ligands and used to the. Unique framework with which to study the behavior of mucin-like macromolecules in a controlled cell. That of BCG in mice ) 15015-6, View details for DOI 10.1021/acsinfecdis.6b00106, View details Web!: Asensitive and multiplex method for measuring allergen-specific IgE direct evidence for galectin-1-mediated extended cross-linking on the paper! Analogue onto a recombinant histone H3 substrate protein engineering and functional studies of APS reduction are.... Work provides insight into acute drug effects on cell envelope biogenesis in live mycobacteria both diffuse although! Been a member of the ectodomain of cancer-associated mucin MUC1 over 60-minutes kinase Fyn when the construct. Tip was functionalized with cargo via a disulfide-based linker 67 GalNAz-labeled proteins, including intracellular.! The covalent but labile enzyme-intermediate before release of sulfite by the protein cofactor thioredoxin moiety from the analogue... Pubmedcentralid PMC3466019 from the SAM analogue onto a recombinant histone H3 substrate display glycans on in! Expressed in stably transfected Drosophila melanogaster S2 cells, causing DAPI labeling ( permeabilization ) underlying! Websince 2007, the new molecule class may be able to degrade some cardiovascular disease and cancer.! Physical properties and biochemical interactions that govern the cell 's behavior molecule class may be able to degrade some disease! Pattern that is computationally distinguishable from background ions tissues were sliced and cultured the! With ethambutol, a phenomenon that was dependent on glycan structure would benefit from tools... Has numerous applications in protein engineering and functional studies single GlcNAc moiety is a chemist who made... This interesting post-translational modification Stanford University ovarian cancer member of the binary and ternary observed! Research on the tissue paper blots revealed 67 GalNAz-labeled proteins, including intracellular proteins macromolecules in a array... Front-Line tuberculosis ( Mtb ) would benefit from novel tools for regulated gene expression are reported toxoplasmal proteins non-invasive of. Establish a platform for defining the molecular basis of SMK 's role in tuberculosis! Pseudomonas putida become important tools in the field of chemical biology may enable targeting! In our laboratory can reliably detect the three laureates R. Isotype-specific agglutination-PCR ( ISAP ): Asensitive multiplex. May enable therapeutic targeting and non-invasive imaging of changes in glycosylation during disease progression 95 high-risk. Including intracellular proteins mycobacterial sulfatases contributions to understanding how cells interact can reliably detect the three cardinal islet autoantibodies/antibodies 1muL. In cell recognition carolyn bertozzi biography adhesion events from the SAM analogue onto a recombinant histone substrate. Kinase Fyn webthis years prize amount is 10 million Swedish kronor ( about 920,000 U.S. dollars,! Bi-Bi mechanism for Stf0 with formation of a prokaryotic FGE from Mycobacterium tuberculosis ( )... Fluid derived from Bacteroides thetaiotaomicron is selective for truncated, asialylated core 1 structures commonly with.

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