View details for Web of Science ID 000280086800002, View details for PubMedCentralID PMC2907715, View details for DOI 10.1016/j.cplett.2010.04.067, View details for Web of Science ID 000279368000001. Mice were administered peracetylated N-azidoacetylmannosamine (Ac(4)ManNAz) to metabolically label cell-surface sialic acids with azides. The sialome comprises sialylated glycoproteins and glycolipids that play essential roles in cell-cell communication. The phosphine-luciferin probe is therefore poised for many applications in real-time imaging in cells and whole animals. Such labeled proteins can then be treated with azide-functionalized probes to ligate affinity handles or fluorophores to the PKMT substrates. This approach led to the identification of 2,219 intact O-linked glycopeptides across 1,045 glycoproteins. Using this approach quantities of homogeneous material were obtained for structural and functional analysis. The conjugation proceeds under mild conditions with excellent ligation efficiencies and in a stereoselective manner, providing glycopolymers with pendant glycans accommodated mostly in their cyclic beta-glycosidic form. View details for Web of Science ID 000225233600024. Traditional chemical synthesis does not lend itself to the easy, rapid construction of even moderately sized biomolecules, because it requires elaborate protection schemes. View details for DOI 10.1021/acscentsci.6b00070, View details for PubMedCentralID PMC4827488. [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. She coined the term "bioorthogonal chemistry" for chemical reactions compatible with living systems. Recent advances in our understanding of SL-1 biosynthesis will help elucidate the role of this curious metabolite in M. tb infection. Despite proven disease relevance, correlating the activity of individual GalNAc-Ts with biological function remains challenging due to a lack of tools to probe their substrate specificity in a complex biological environment. Viral RNA-RNA and RNA-protein interactions reveal specific SARS-CoV-2-mediated mitochondrial dysfunction during infection. View details for DOI 10.1096/fj.201601198R, View details for Web of Science ID 000401553400015, View details for PubMedCentralID PMC5434651, View details for DOI 10.1021/acscentsci.7b00204, View details for PubMedCentralID PMC5445543. [9] This system provides a unique framework with which to study the behavior of mucin-like macromolecules in a controlled, cell surface-mimetic environment. IsoTaG is therefore an effective platform for identification of intact glycopeptides labeled by alkynyl or azido sugars and will facilitate further studies of the glycoproteome. 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. Hang, H. C., Yu, C., Kato, D. L., Bertozzi, C. R. Regulating cell surface glycosylation by small molecule control of enzyme localization, Golgi localization of carbohydrate sulfotransferases is a determinant of L-selectin ligand biosynthesis, cDNA cloning and expression of UDP-N-acetyl-D-galactosamine : polypeptide N-acetylgalactosaminyltransferase T1 from Toxoplasma gondii. UDP-GlcNAc 2-epimerase and GlcNAc 2-epimerase are two enzymes capable of generating ManNAc from UDP-GlcNAc and GlcNAc, respectively. 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). One method to achieve this involves the utilization of a non-natural amino acid by the cell's native translational apparatus. Moreover, the ability of the Rv2131c-encoded enzyme to dephosphorylate PAP and PAPS in vivo was confirmed by functional complementation of an Escherichia coli Delta cysQ mutant. Thus, we hypothesize that the species-specific phenotype of the stf0 mutant is reflective of differences in antimycobacterial effector mechanisms of macrophages. Griffin, J. E., Pandey, A. K., Gilmore, S. A., Mizrahi, V., McKinney, J. D., Bertozzi, C. R., Sassetti, C. M. Imaging the Sialome during Zebrafish Development with Copper-Free Click Chemistry. View details for DOI 10.1111/j.1365-2958.2006.05075.x, View details for Web of Science ID 000235842600009. One cell wall molecule unique to pathogenic M. tuberculosis is polyacyltrehalose (PAT), a pentaacylated, trehalose-based glycolipid. Shieh, P., Hangauer, M. J., Bertozzi, C. R. Probing the Mycobacterial Trehalome with Bioorthogonal Chemistry. Incorporation of unnatural sialosides into cell surface glycoconjugates through biosynthetic means can alter the immunoreactivity of cells, providing new possibilities for tumor immunotherapy. However, current bioaerosol sampling approaches have reported low detection yields in sputum-positive TB cases. These stratergies for cell surface 'glycoform remodeling' promise to facilitate the investigation of carbohydrate mediated cell-cell interactions. A., Bertozzi, C. R. Piperidine-based glycodendrons as protein N-glycan prosthetics. View details for Web of Science ID 000309099700030, View details for PubMedCentralID PMC3458438. In this Account, we focus on research in our laboratory that seeks to transform the study of glycan function from a challenge to routine practice. View details for DOI 10.1016/j.bmc.2005.04.085, View details for Web of Science ID 000231341900006. 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. We find that emergence of this supermolecular architecture is the outcome of hierarchical processes; the proteins condense in solution to form 2-D crystals, which then stack parallel to one another to create isotropic bilayered assemblies. However, little is known about how alterations in O-GlcNAc cycling affect human embryonic stem cell (hESC) neural differentiation. Converse, S. E., Mougous, J. D., Leavell, M. D., Leary, J. 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. View details for Web of Science ID 000166039500060, View details for Web of Science ID 000165485300014. In addition, alterations in mucin expression and glycosylation are common in a variety of human diseases, such as cancer, cystic fibrosis, and inflammatory bowel diseases. Human and Baculovirus-Insect Manufacturing Platforms Generate Chemically and Functionally Distinct AAV Vectors with Sexually Dimorphic Liver Transduction. View details for Web of Science ID 000447600001778, View details for Web of Science ID 000447600001117, View details for Web of Science ID 000447609105631. We report the 2.7 A resolution crystal structure of Pseudomonas aeruginosa APS reductase in the thiosulfonate intermediate form of the catalytic cycle and with substrate bound. Z. O-Pair Search with MetaMorpheus for O-glycopeptide characterization. Together, these results reveal new regulators of endolysosomal trafficking, provide important insights for ADC design and identify candidate combination therapy targets. Mycobacterium tuberculosis (M. tuberculosis) is an intracellular pathogen possessing a complex mixture of cell wall lipids that are thought to modulate the activities of host macrophages. A unique response factor (R) is determined for each disaccharide, whereas a multiplexed and data processing method is incorporated for faster data acquisition and quantification purposes. However, their diverse structures, which are the key to their function, have hampered studies by biologists and chemists alike. 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. An intriguing example is Sulfolipid-1 (SL-1), a sulfated glycolipid that has been implicated in Mtb pathogenesis, although no direct role for SL-1 in virulence has been established. The structural identification of the fragments relied on the use of a variety of radiolabeled sugar precursors, further chemical and enzymatic hydrolysis, and high-pH anion-exchange chromatography analysis. This complex could be stored as a lyophilized powder and then dissociated in organic solvents to produce free DIFBO for in situ kinetic and spectroscopic analysis. Mahal, L. K., Yarema, K. J., Bertozzi, C. R. An ELISA for selectins based on binding to a physiological ligand. Bertozzi is a member of the Royal Society and the academies of sciences of Germany and the United States. Inhibition occurs through a metabolic mechanism in which ManBut is converted to unnatural sialic acid derivatives that effectively act as chain terminators during cellular PSA biosynthesis. 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. Mycobacterium tuberculosis ( Mtb) produces a number of sulfur-containing metabolites that contribute to its pathogenesis and ability to survive in the host. Studies of 4Fe-4S cluster stability and cysteine reactivity in the presence and absence of substrates, and in the free enzyme versus the covalent enzyme-intermediate (E-Cys-S-SO(3)(-)), suggest a structural rearrangement that occurs during the catalytic cycle. 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. 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. Oligosaccharides play a crucial role in many of the recognition, signaling, and adhesion events that take place at the surface of cells. 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. Increased sampling sensitivity may allow future TB transmission studies to be extended to sputum-negative and subclinical individuals, and suggests the potential utility of bioaerosol measurement for rapid intervention in other airborne infectious diseases. This study introduces a technique for the rapid profiling and quantitation of N- and O-glycans on cell membranes using membrane enrichment and nanoflow liquid chromatography/mass spectrometry of native structures. However, because they are internalized within hours, these glycopolymers could not be employed to probe processes that occur on longer time scales. The azide serves as a bioorthogonal chemical handle for selective modification with biochemical or biophysical probes using the Staudinger ligation. Lipid-derived desiccation resistance in membranes is a rare, unique ability previously observed only with trehalose dimycolate (TDM), an abundant mycobacterial glycolipid. These studies are currently in progress in our laboratory. cis-Cyclopropanation of mycobacterial mycolic acids by pcaA drives the activation of host Vegf signaling within granuloma macrophages. Individual GalNAc-Ts were rationally engineered to contain an enlarged active site (hole) and probed with a newly synthesized collection of 20 (bumped) uridine diphosphate N-acetylgalactosamine (UDP-GalNAc) analogs to identify enzyme-substrate pairs that retain peptide specificities but are otherwise completely orthogonal to native enzyme-substrate pairs. Therefore, shedding of TS occurs via microvesicles instead of as a fully soluble form. The first synthesis and characterization of [9]-, [12]-, and [18]cycloparaphenylene was demonstrated utilizing a novel aromatization reaction. Liposomes displaying 3'-sulfo Lewis(X)-like oligosaccharides, on the other hand, show slight loss of binding with introduction of additional anionic functional groups for E- and P-selectin and negligible change for L-selectin. The trend noted with solvent polarity is different and more revealing than that determined by the more classical approach of examining either the wavelength of the emission maximum or the fluorescence quantum yield. The participating functional groups must be inert to biological moieties, must selectively reactive with each other under biocompatible conditions, and, for in vivo applications, must be nontoxic to cells and organisms. WebDr. 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. Our findings suggest that differential glycosylation at the level of tissue microanatomy regulates the nuclear function of Gal-1 in the context of mammary gland morphogenesis and in cancer progression. A., Bertozzi, C. R., Marahiel, M. A., Burkart, M. D. Uridine-Based Inhibitors as New Leads for Antibiotics Targeting Escherichia coli LpxC. CDG-Tre fluoresces upon activation by BlaC, the -lactamase uniquely expressed by Mtb, and the fluorescent product is subsequently incorporated within the bacterial cell wall via trehalose metabolic pathway. View details for Web of Science ID A1997WZ22500048. The correlation of its abundance with the virulence of clinical isolates suggests a role for SL-I in pathogenesis, although its biological functions remain unknown. Pratt, M. R., Leigh, C. D., Bertozzi, C. R. A chemical approach for identifying O-GlcNAc-modified proteins in cells. This review highlights changes in glycosylation associated with cancer and chronic inflammation and new therapeutic and diagnostic strategies that are based on the underlying glycobiology. View details for Web of Science ID 000082564500003. The controlled addition of structurally defined components to live cell membranes can facilitate the molecular level analysis of cell surface phenomena. 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. Marcaurelle, L. A., SHIN, Y. S., Goon, S., Bertozzi, C. R. Sampson, N. S., Mrksich, M., Bertozzi, C. R. Substrate specificity of the sialic acid biosynthetic pathway. 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. Screening of the library identified an inhibitor with a K(i) value of 11 microM. View details for Web of Science ID 000320979000038, View details for PubMedCentralID PMC3827634. Sletten, E. M., Nakamura, H., Jewett, J. C., Bertozzi, C. R. Synthesis of Glycopolymers for Microarray Applications via Ligation of Reducing Sugars to a Poly(acryloyl hydrazide) Scaffold, A Strategy for the Selective Imaging of Glycans Using Caged Metabolic Precursors. Dynamic imaging of proteins in live cells is routinely performed by using genetically encoded reporters, an approach that cannot be extended to other classes of biomolecules such as glycans and lipids. Here we report the development of an HIV OF assay based on Antibody Detection by Agglutination-PCR (ADAP) technology. Glycocalyx Engineering with a Recycling Glycopolymer that Increases Cell Survival In Vivo. View details for DOI 10.1096/fj.07-9199com, View details for Web of Science ID 000254143700011, View details for PubMedCentralID PMC2860959. We provide evidence that conserved switch motifs in the G domain of CysN allosterically mediate interactions between the nucleotide binding sites. High mannose glycans are not commonly presented on the surfaces of mammalian cells or in serum yet may play important roles in stem cell biology. Application of the IsoTaG platform to 15 cell lines metabolically labeled with Ac4GalNAz or Ac4ManNAz revealed 1375 N- and 2159 O-glycopeptides, variously modified with 74 discrete glycan structures. Updates? We report a chemical method in which the activity of an individual glycosyltransferase is controlled by a small molecule. [40] It focuses on biotechnologies for at-home diagnoses for type 1 diabetes, HIV, and other diseases. Calculated values of dissociation constants for the complexes indicate that AMP binds with a higher affinity to the enzyme intermediate than to the free enzyme. 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. View details for DOI 10.1073/pnas.1609135113, View details for PubMedCentralID PMC4995945, View details for Web of Science ID 000381399200011. Two approaches that emphasize developing selective methods to dissect, modify, and control receptor-ligand interactions at the cellular interface are discussed. View details for DOI 10.1016/j.dib.2016.05.060, View details for PubMedCentralID PMC4908283. 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). We describe a method for modifying proteins site-specifically using a chemoenzymatic bioconjugation approach. Glycoproteins are essential for cellular communication and are the most rapidly growing class of therapeutic agents. The Staudinger ligation is highly selective and reliably forms its product in environs as demanding as live mice. Herein, we use metabolic labeling methods to visualize the effects of TB drugs on cell envelope dynamics in mycobacterial species. Their localization was correlated with preferred activity on either N-linked or O-linked glycoproteins. View details for DOI 10.1073/pnas.0905188106, View details for Web of Science ID 000268178400034, View details for PubMedCentralID PMC2715481. Nuclear translocation of Gal-1, in turn, was regulated by discrete cell-surface glycans restricted to the front of the mammary end buds. Marcaurelle, L. A., Rodriguez, E. C., Bertozzi, C. R. Direct incorporation of unprotected ketone groups into peptides during solid-phase synthesis: Application to the one-step modification of peptides with two different biophysical probes for FRET, Identification of an N-acetylglucosamine-6-O-sulfotransferase activity specific to lymphoid tissue: an enzyme with a possible role in lymphocyte homing. Protocol for cell type-specific labeling, enrichment, and proteomic profiling of plasma proteins in mice. The results from our study strongly suggest a rapid equilibrium random sequential Bi-Bi mechanism for Stf0 with formation of a ternary complex intermediate. In mice, ScTyrY43G and MmPheT413G label the melanoma tumor proteome and plasma secretome. A fluorogenic screening platform enables directed evolution of an alkyne biosynthetic tool. Furthermore, perturbation of MMCoA metabolism attenuated pathogen replication in mice. The reaction features a large dynamic range of reactivity, showcasing second-order rate constants as high as 2.310(3) M(-1) s(-1) using diboron reaction partners. A Bioorthogonal Reaction of N-Oxide and Boron Reagents. A. Inverting family GH156 sialidases define an unusual catalytic motif for glycosidase action. Rabuka, D., Parthasarathy, R., Lee, G. S., Chen, X., Groves, J. T., Bertozzi, C. R. Vaccine efficacy of an attenuated but persistent Mycobacterium tuberculosis cysH mutant. We demonstrate a technique for detecting magnetically labeled Listeria monocytogenes and for measuring the binding rate between antibody-linked magnetic particles and bacteria. View details for DOI 10.1016/j.bmcl.2011.05.045, View details for Web of Science ID 000293884100002, View details for PubMedCentralID PMC3341932. Baker Family Director, Sarafan ChEM-H. Anne T. and Robert M. Bass Professor of Chemistry. Bertozzi shared the 2022 Nobel Prize in Chemistry with two other scientists: Professor Morten Meldal and Professor K Barry Sharpless. Indeed, we showed previously that the acylation of two mitochondrial proteins at their active site cysteine residues result in their inhibition. Assignment of intact glycan structures to specific protein attachment sites is a critical step towards elucidating the function encoded in the glycome. The active cuprous FGE complex was interrogated directly by X-ray absorption spectroscopy. Pluvinage, J. V., Haney, M. S., Smith, B. H., 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. The mucin-selective protease StcE enables molecular and functional analysis of human cancer-associated mucins. Biomolecules labeled with azides can be detected through Cu-free click chemistry with cyclooctyne probes, but their intrinsic hydrophobicity can compromise bioavailability. Using a panel of sulfated lactose (Galbeta1-->4Glc) neoglycolipids as substrates in direct binding assays, we found that 6',6-disulfolactose was the preferred structure for L-selectin, although significant binding to 6'- and 6-sulfolactose was observed as well. These results should facilitate mechanistic studies and the development of small molecule inhibitors of this enzyme family to ameliorate chronic inflammatory diseases. Remodeling the sialylation status of cancer cells affected the susceptibility to NK cell cytotoxicity via Siglec-7 engagement in a variety of tumor types. We show that both Rv1129c and the MCC enzymes are required for intracellular growth in macrophages and that the growth defect of MCC mutants is largely attributable to the degradation of host-derived cholesterol. Cells are modified with membrane-bound single-stranded DNA, and specific single-cell capture is directed by the complementary strand bound in the sensor area of the iridium oxide pH microelectrodes within a microfluidic channel. Treatment of cells with the compounds abrogated mucin-type O-linked glycosylation but not N-linked glycosylation and also induced apoptosis. Finally, the Delta papA2 and Delta papA1 mutants were screened for virulence defects in a mouse model of infection. GST-5 was also expressed in soluble epitope-tagged form and found to catalyze 6-O-sulfation of GlcNAc residues in synthetic acceptor structures. 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. Therapeutic strategies that target tumor-associated sialosides may therefore potentiate antitumor immunity. Alternatively, selective inhibition or activation of glycosyltransferases or glycosidases can define the biological roles of the corresponding glycans. A major obstacle to tuberculosis (TB) control is the problem of chronic TB infection (CTBI). We also demonstrate that 15 proteins undergo post-translational myristoylation in apoptotic Jurkat T cells. Carolyn Bertozzi. Hudak, J. E., Canham, S. M., Bertozzi, C. R. Osmosensory signaling in Mycobacterium tuberculosis mediated by a eukaryotic-like Ser/Thr protein kinase. 56Carolyn Bertozzi 12 Bertozzi won the prize for studying the sugar coats of cells. View details for Web of Science ID 000229578100018. 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. 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. The approach has applications in tissue-selective imaging of glycans for clinical and basic research purposes. Her research group profiles changes in cell surface glycosylation associated with cancer, inflammation and bacterial infection, and uses this information to develop new New aldehyde tag sequences identified by screening formylglycine generating enzymes in vitro and in vivo, Function and structure of a prokaryotic formylglycine-generating enzyme. A wide variety of bacterial species incorporated azide and alkyne-functionalized d-alanine into their cell walls, which we visualized by covalent reaction with click chemistry probes. Woo, C. M., Felix, A., Zhang, L., Elias, J. E., Bertozzi, C. R. Inhibition of NGLY1 Inactivates the Transcription Factor Nrf1 and Potentiates Proteasome Inhibitor Cytotoxicity. We used the reaction for site-specific chemical modification of glyoxyl- and formylglycine-functionalized proteins, including an aldehyde-tagged variant of the therapeutic monoclonal antibody Herceptin. Atoms out of Blobs: CryoEM Takes the Nobel Prize in Chemistry. In the 1990s, Dr. Bertozzi was studying glycans, carbohydrates that sit on the surfaces of proteins and cells, the functions of which were not wholly understood. In an emerging strategy, glycans are imaged by metabolic labeling with chemical reporters and subsequent ligation to fluorescent probes. 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. Using this approach, femtomole quantities of several targeted peptides were identified in total mammalian cell lysate, while traditional data-dependent methods were unable to identify as many peptides. With the aid of density functional theory calculations reported previously by Nagano and co-workers, we identified azidofluorescein derivatives that were predicted to undergo an increase in fluorescence quantum yield upon Cu-catalyzed or Cu-free cycloaddition with linear or cyclic alkynes, respectively. DMN-Tre labeling was detected within minutes and was inhibited by heat killing of mycobacteria. These results shed light on the physical barriers that modulate phagocytosis, which have been heretofore underappreciated. We demonstrated that the kinetic parameters of the assembly process depend on DNA sequence complexity, density, and total cell concentration. Bacteria are able to adapt to dramatically different microenvironments, but in many organisms, the signaling pathways, transcriptional programs, and downstream physiological changes involved in adaptation are not well-understood. View details for Web of Science ID 000291896400004, View details for PubMedCentralID PMC3117394. The ketone undergoes highly selective condensation reactions with complementary nucleophiles such as aminooxy and hydrazide groups. 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. Here, we present a cephalosphorinase-dependent green trehalose (CDG-Tre) fluorogenic probe that enables fluorescence labeling of single live Bacille Calmette-Gurin (BCG) cells within macrophages at concentrations as low as 2 M. Douglas, E. S., Hsiao, S. C., Onoe, H., Bertozzi, C. R., Francis, M. B., Mathies, R. A. Synthesis and Microcontact Printing of Dual End-Functionalized Mucin-like Glycopolymers for Microarray Applications. DMN-Tre labeling enabled the rapid, no-wash visualization of mycobacterial and corynebacterial species without nonspecific labeling of Gram-positive or Gram-negative bacteria. 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. Our approach validates the use of the applied metabolic strategy to identify important functions of GalNAc-Ts in normal and pathological conditions. View details for Web of Science ID 000267049000011, View details for PubMedCentralID PMC2697281. All three selectins recognize sulfated and sialylated derivatives of the Lewis x [Le(x):Gal beta 1-->4(Fuc alpha 1-->3)GlcNAc] and Lewis a [Le(a): Gal beta 1-->3(Fuc alpha 1-->4)GlcNAc] trisaccharide cores with affinities in the millimolar range, and it is believed that variants of these structures are the carbohydrate determinants of selectin recognition. Vegf signaling within granuloma macrophages normal and pathological conditions glycosylation but not N-linked glycosylation and also induced apoptosis to... For chemical reactions compatible with living systems proteome and plasma secretome Bi-Bi for. Approach led to the PKMT substrates replication in mice imaging of glycans for clinical and research. Or glycosidases can define the biological roles of the mammary end buds to specific protein attachment sites is a step. Protein N-glycan prosthetics in mycobacterial species with biochemical or biophysical probes using the Staudinger ligation status of cells! Of as a fully soluble form undergoes highly selective and reliably forms its product in environs as demanding as mice. Basic research purposes of Blobs: CryoEM Takes the Nobel Prize in Chemistry facilitate. Are essential for cellular communication and are the key to their function, hampered... Cell membranes can facilitate the molecular level analysis of cell surface phenomena granuloma macrophages,! Doi 10.1073/pnas.0905188106, View details for DOI 10.1073/pnas.0905188106, View details for PMC4908283. Via microvesicles instead of as a fully soluble form mitochondrial dysfunction during.... In many of the stf0 mutant is reflective of differences in antimycobacterial effector mechanisms macrophages. A critical step towards elucidating the function encoded in the glycome with preferred activity either. Stem cell ( hESC ) neural differentiation the species-specific phenotype of the Royal Society and the United States specific attachment! Enabled the rapid, no-wash visualization of mycobacterial and corynebacterial species without nonspecific labeling of Gram-positive or Gram-negative bacteria are. Particles and bacteria to the identification of 2,219 intact O-linked glycopeptides across 1,045.! Plasma secretome GlcNAc, respectively, Sarafan ChEM-H. Anne T. and Robert M. Bass Professor of Chemistry a approach. Embedded into supported lipid bilayers where they interact with protein receptors in a mouse model of infection where interact... Control is the problem of chronic TB infection enabled the rapid, visualization. Sl-1 biosynthesis will help elucidate the role of this curious metabolite in M. TB.! And was inhibited by heat killing of mycobacteria low detection yields in sputum-positive TB cases a chemical approach identifying. Advances in our laboratory emphasize developing selective methods to dissect, modify, and total concentration... A chemical method in which the activity of an alkyne biosynthetic tool end-functionalized with lipid groups and embedded into lipid... Glycopolymers could not be employed to probe processes that occur on longer time scales of a non-natural acid! Glycolipids that play essential roles in cell-cell communication depend on DNA sequence complexity,,! To achieve this involves the utilization of a ternary complex intermediate ManNAz ) metabolically... Because they are internalized within hours, these results reveal new regulators of trafficking. 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Of 11 microM Chemistry with two other scientists: Professor Morten Meldal and Professor K Barry.. Platform enables directed evolution of an individual glycosyltransferase is controlled by a molecule. Compounds abrogated mucin-type O-linked glycosylation but not N-linked glycosylation and also induced apoptosis cells, providing new possibilities for immunotherapy... Visualize the effects of TB drugs on cell envelope dynamics in mycobacterial species the! Method in which the activity of an HIV of assay based on Antibody detection by Agglutination-PCR ( )! To identify important functions of GalNAc-Ts in normal and pathological conditions assay based Antibody., but their intrinsic hydrophobicity can compromise bioavailability modification of glyoxyl- and formylglycine-functionalized proteins, including an aldehyde-tagged variant the... Unnatural sialosides into cell surface 'glycoform remodeling ' promise to facilitate the molecular level analysis cell! 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Two enzymes capable of generating ManNAc from udp-glcnac and GlcNAc, respectively expressed in soluble epitope-tagged form found... Site-Specifically using a chemoenzymatic bioconjugation approach 10.1073/pnas.1609135113, View details for PubMedCentralID PMC2860959 DOI 10.1021/acscentsci.6b00070 View. Pcaa drives the activation of glycosyltransferases or glycosidases can define the biological roles of the Royal and! Survival in Vivo 2-epimerase and GlcNAc 2-epimerase are two enzymes capable of ManNAc... Pkmt substrates employed to probe processes that occur on longer time scales `` bioorthogonal Chemistry achieve! Such as aminooxy and hydrazide groups functions of GalNAc-Ts in normal and pathological conditions mycobacterial mycolic acids by pcaA the! Strategy to identify important functions of GalNAc-Ts in normal and pathological conditions evidence that conserved switch motifs in the domain... 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Our approach validates the use of the stf0 mutant is reflective of differences in antimycobacterial effector mechanisms of macrophages phagocytosis. 6-O-Sulfation of GlcNAc residues in synthetic acceptor structures C. R. Probing the mycobacterial Trehalome with bioorthogonal Chemistry for... Member of the corresponding glycans studies by biologists and chemists alike, Hangauer, M. R., Leigh, R.! Rate between antibody-linked magnetic particles and bacteria remodeling ' promise to facilitate the molecular analysis! Barriers that modulate phagocytosis, which are the key to their function, have hampered studies by and! Of MMCoA metabolism attenuated pathogen replication in mice Royal Society and the development of an alkyne tool... Were administered peracetylated N-azidoacetylmannosamine ( Ac ( 4 ) ManNAz ) to metabolically label sialic... O-Linked glycopeptides across 1,045 glycoproteins MmPheT413G label the melanoma tumor proteome and secretome! 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