As a proof-of-concept, we have used SETDB1 to transfer the alkyne moiety from the SAM analogue onto a recombinant histone H3 substrate. Palaniappan, K. K., Hangauer, M. J., Smith, T. J., Smart, B. P., Pitcher, A. However, for targeting of hydrophobic fluorophores such as ATTO 647N, the hydrophobicity of ADIBO was detrimental, and superior targeting was achieved by conjugation to the less hydrophobic monofluorinated cyclooctyne (MOFO). Growth then proceeds by formation of new tetramers exclusively at cluster edges, implying tetramer formation is autocatalytic. Phosphatidylinositol-phospholipase-C activity is actually not present in trypomastigotes. Taken together, these studies indicate that Rv2131c encodes a CysQ enzyme that may play a role in mycobacterial sulfur metabolism. This novel assay may improve pediatric testing compliance and facilitate easier community-wide screening for islet autoantibodies. A., Baskin, J. M., Bertozzi, C. R., Koberstein, J. T., Turro, N. J. We employed BARAC for live cell fluorescence imaging of azide-labeled glycans. Treatment of cells with the compounds abrogated mucin-type O-linked glycosylation but not N-linked glycosylation and also induced apoptosis. Highlights of recent progress include an extension of the list of instances of selectin participation in inflammatory diseases, further definition of selectin carbohydrate specificities, and identification of their carbohydrate-based ligands. 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. Mycobacteria, including the pathogen Mycobacterium tuberculosis, use the non-mammalian disaccharide trehalose as a precursor for essential cell-wall glycolipids and other metabolites. Higher order oligosaccharides were readily generated by alkylation of the corresponding 3-thioGalNAc with N-bromoacetamido sugars. Grand Challenges in Chemistry for 2016 and Beyond. In addition, we generated a mutant M. tuberculosis strain lacking FGE. Positioned at the C-terminus of many eukaryotic proteins, the glycosylphosphatidylinositol (GPI) anchor is a posttranslational modification that anchors the modified protein in the outer leaflet of the cell membrane. A goal of tissue engineering is to produce a scaffold material that will guide cells to differentiate and regenerate functional replacement tissue at the site of injury. These uridine analogs represent the first generation of chemical tools to study the functions of mucin-type O-linked glycosylation. Using high-performance liquid chromatography, we quantified the degree of accumulation and reversibility upon acidic compartment neutralization in macrophages and observed that accumulation was greater in infected than in uninfected macrophages. The kinetics of this reaction are of paramount importance for studies of dynamic processes, particularly in living subjects. Aldehyde- and ketone-functionalized proteins are appealing substrates for the development of chemically modified biotherapeutics and protein-based materials. View details for Web of Science ID 000275868700024, View details for PubMedCentralID PMC2840677. Collectively, these chemical approaches are contributing great insight into the myriad biological functions of oligosaccharides. WebCarolyn Ruth Bertozzi (born October 10, 1966) is an American chemist and Nobel laureate, known for her wide-ranging work spanning both chemistry and biology. View details for DOI 10.1074/jbc.M204613200, View details for Web of Science ID 000177859000029. SL-1 consists of a trehalose-2-sulfate (T2S) disaccharide elaborated with four lipids. Notably, PapA3 was selective for trehalose; no activity was observed with other structurally related disaccharides. We demonstrate here that a previously uncharacterized sulfated molecule, termed S881, is localized to the outer envelope of M. tuberculosis and negatively regulates the virulence of the organism in two mouse infection models. Harland, C. W., Rabuka, D., Bertozzi, C. R., Parthasarathy, R. Rv2131c from Mycobacterium tuberculosis is a CysQ 3 '-phosphoadenosine-5 '-phosphatase. This strategy mimics the natural glycosylphosphatidylinositol (GPI) linkage found in many natural membrane-associated proteins; however, the synthetic method utilizes simple lipid anchors without glycans. Sulfated constituents of GlyCAM-1 were identified as Gal-6-SO4, GlcNAc-6-SO4, (SO4-6)Gal beta 1-->4GlcNAc, and Gal beta 1-->4(SO4-6)GlcNAc. 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. [6] In 1999, while working with HHMI and at Berkeley, she founded the field of bioorthogonal chemistry and coined the term in 2003. View details for DOI 10.1074/jbc.M304928200, View details for Web of Science ID 000185713800121. This system provides a unique framework with which to study the behavior of mucin-like macromolecules in a controlled, cell surface-mimetic environment. Antigen presentation to Tcells in major histocompatibility complex class II (MHC class II) requires the conversion of early endo/phagosomes into lysosomes by a process called maturation. In this study, we examine the nature of the sulfate-modified carbohydrates of GlyCAM-1. Using a recently described method (Mahal, L. K., Yarema, K. J., and Bertozzi, C. R. (1997) Science 276, 1125-1128), we delivered a uniquely reactive ketone group to endogenous cell surface sialic acid residues by treating cells with the ketone-bearing metabolic precursor N-levulinoylmannosamine (ManLev). Monomeric sialyl Lewis(X) (sLe(x)) and sLe(x)-like oligosaccharides are minimal structures capable of supporting selectin binding in vitro. Consistent with other studies, we find that O-GlcNAc sites in T cells lack a strict consensus sequence. Bertozzi developed a new type of reaction, now called a bioorthogonal reaction, that can occur in living environmentsrather than just in labswith no interference with other biochemical processes. This data also suggested for a role of GALNT3 in aberrant EOC glycosylation, possibly implicated in disease progression. [reaction: see text], View details for Web of Science ID 000174997600030. View details for Web of Science ID 000263320900008, View details for PubMedCentralID PMC2709987. A., Hangauer, M. J., Bertozzi, C. R. PapA1and PapA2 are acyltransferases essential for the biosynthesis of the Mycobacterium tuberculosis virulence factor Sulfolipid-1. In an effort to mimic the high-affinity binding, polyvalent scaffolds that contain multicomponent displays of selectin-binding ligands have been synthesized. The dependence of the desiccation protection on the synthetic trehalose glycolipid fraction is nearly identical to that of TDM. Palaniappan, K. K., Pitcher, A. Mucin-type O-linked glycosylation is a fundamental post-translational modification that is involved in a variety of important biological processes. The use of symbols to depict glycans originated from Kornfeld in 1978, was systematized in the First Edition of "Essentials of Glycobiology" and updated for the second edition, with input from relevant organizations such as the Consortium for Functional Glycomics. However, when activity of the vacuolar H+-ATPase was also inhibited, disulfide reduction decreased SHGFP-MUC5AC/CK t((1/2)) while diminishing its intraluminal concentration. The approach has applications in tissue-selective imaging of glycans for clinical and basic research purposes. View details for DOI 10.1128/AAC.01639-16, View details for PubMedCentralID PMC5328571. Glycocalyx Engineering with a Recycling Glycopolymer that Increases Cell Survival In Vivo. Here we report a method for rapid profiling of fucosylated glycoproteins from human cells using 6-azido fucose as a metabolic label. With this technique, we studied the dynamics of glycan trafficking and identified a population of sialoglycoconjugates with unexpectedly rapid internalization kinetics. Riley, N. M., Lu, L., Shortreed, M. R., Smith, L. M., Bertozzi, C. R. Dissecting O-GalNAc glycosylation by glycosyltransferase engineering. View details for Web of Science ID 000081721100060, View details for Web of Science ID 000080303100023, View details for Web of Science ID 000080212600003, View details for Web of Science ID 000079041700025. Previously, we described the biochemical activity of the sulfotransferase Stf0 that initiates SL-1 biosynthesis. View details for Web of Science ID 000232605600062. Acute Modulation of Mycobacterial Cell Envelope Biogenesis by Front-Line Tuberculosis Drugs. View details for Web of Science ID 000275864500006, View details for PubMedCentralID PMC2865253. Carolyn Ruth Bertozzi (born October 10, 1966) is an American chemist and Nobel laureate, known for her wide-ranging work spanning both chemistry and biology. Both normal and cancerous prostate tissues were sliced and cultured in the presence of the azide-functionalized sialic acid biosynthetic precursor Ac4 ManNAz. 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. Using a high-throughput enzyme-linked lectin assay (ELLA), two inhibitors of murine ppGalNAcT-1 (K(I) approximately 8 microM) were identified that also inhibit several other members of the family. Upon changing to differentiation media, myotubes formed in the center of the patterned areas with an excellent degree of edge alignment. View details for Web of Science ID 000077466300003, View details for Web of Science ID 000077383600001. The Staudinger ligation takes advantage of the electrophilicity of the azide; however, the azide can also participate in cycloaddition reactions. A correlation between hypersialylation and immunoprotection has been observed, but few hypotheses have provided a mechanistic understanding of this immunosuppressive phenomenon. Shurer, C. R., Kuo, J., Roberts, L., Gandhi, J. G., Colville, M. J., Enoki, T. A., Pan, H., Su, J., Noble, J. M., Hollander, M. J., O'Donnell, J. P., Yin, R., Pedram, K., Mockl, L., Kourkoutis, L. F., Moerner, W. E., Bertozzi, C. R., Feigenson, G. W., Reesink, H. L., Paszek, M. J. 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. Our results suggest a correlation between decreased alkyne bond angle and increased cyclooctyne reactivity. The sialome comprises sialylated glycoproteins and glycolipids that play essential roles in cell-cell communication. 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. 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. Cambier, C. J., Banik, S. M., Buonomo, J. Here we describe the identification of a prokaryotic FGE from Mycobacterium tuberculosis. Both the Staudinger ligation and the strain-promoted [3 + 2] cycloaddition using optimized cyclooctynes were effective for tagging azides on live cells. Our goal with this survey is to provide a foundation on which continued technological advancements can be made to promote further explorations of protein glycosylation. 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. A., Sletten, E. M., Bertozzi, C. R., Popik, V. V., Ting, A. Y. The exquisite chemical selectivity required of this process is supplied by the Staudinger ligation of azides and phosphines, a reaction that has been performed on cultured cells without detriment to their physiology. In a prototypical experiment, a unique chemical motif, often as small as a single functional group, is incorporated into the target biomolecule using the cell's own biosynthetic machinery. These findings suggest that high mannose glycans are the major component of cell surface glycosylation with even terminal glucoses. View details for DOI 10.1073/pnas.0610634104, View details for Web of Science ID 000245256700066, View details for PubMedCentralID PMC1829275. 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. We also probed the relationship between mycomembrane and peptidoglycan dynamics using a dual metabolic labeling strategy. Tuberculosis (TB) disease is a global epidemic caused by the pathogenic Mycobacterium tuberculosis (Mtb). We performed in vivo reconstitution experiments in which ST8Sia IV(-/-) progenitors competed with wild-type cells to repopulate depleted or deficient immune subsets. Here, we demonstrate that, relative to wild-type controls, ST8Sia IV(-/-) mice have a 30% reduction in total thymocytes and a concomitant deficiency in the earliest thymocyte precursors. Nevertheless, CD45 remained to be the main acceptor. Herein we describe a novel glycosyltransferase assay that exploits their unnatural substrate tolerance and the unique chemical reactivity of the azide. Low molecular weight and singly charged fragments, obtained by a combination of gel filtration and anion-exchange chromatography, were analyzed. View details for Web of Science ID 000224032900044. A chimera comprising the localization domain of GlcNAc6ST-1 fused to the catalytic domain of GlcNAc6ST-2 was confined to the trans-Golgi network and adopted the substrate preference of GlcNAc6ST-1. The objective of these methods is to make glycoconjugate synthesis accessible to a broader community, thereby accelerating progress in glycobiology. Zhou, X., Rodriguez-Rivera, F. P., Lim, H., Bell, J. C., Bernhardt, T. G., Bertozzi, C. R., Theriot, J. A., Bertozzi, C. R. Compositional profiling of heparin/heparan sulfate using mass spectrometry: assay for specificity of a novel extracellular human endosulfatase. 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. To identify a tagged-peptide's sequence, we performed an inclusion list-driven shotgun proteomics experiment where peptides bearing a recoded mass envelope were targeted for fragmentation, allowing for direct site mapping. Redundancy of related family members and embryonic lethality both complicate genetic methods for deconvoluting functions of glycosyltransferases. Long-term central nervous system delivery of an antibody that blocks CD22 function reprograms microglia towards a homeostatic transcriptional state and improves cognitive function in aged mice. Parthasarathy, R., Rabuka, D., Bertozzi, C. R., Groves, J. T. Copper-free click chemistry for dynamic in vivo imaging. This reaction has a second-order rate constant of 0.25 M(-1) s(-1), on par with fast bioorthogonal reactions of azides, and proceeds readily in aqueous environments. Presented herein is the synthesis and evaluation of a bisubstrate analogue designed to inhibit estrogen sulfotransferase. Such metabolic interference can block the expression of oligosaccharides or alter the structures of the sugars presented on cells. This template-driven mineralization technique provides an efficient approach toward bonelike composites with high mineral-hydrogel interfacial adhesion strength. Finally, the lipase inhibitor tetrahydrolipstatin disrupts Chp1 activity in M. tuberculosis, suggesting an avenue for perturbing SL-1 biosynthesis in vivo. A novel germline variant in CSF3R reduces N-glycosylation and exerts potent oncogenic effects in leukemia. View details for DOI 10.1074/jbc.M111.315473, View details for Web of Science ID 000301349400015. We were able to bypass the salvage pathway by using an azide-functionalized analogue of GDP-fucose. She became an assistant professor at Berkeley in 1996 and a full professor of chemistry and molecular and cell biology in 2002. Here, we describe a method for visualizing and analyzing organelle- and stimulus-specific O-GlcNAcylated proteins and use it to identify the mitochondrial voltage-dependent anion channel 2 (VDAC2) as an O-GlcNAc substrate. Ligation of synthetic lipids with designed anchor structures to proteins was performed using native chemical ligation (NCL) of a C-terminal peptide thioester and an N-terminal cysteine lipid. Many cellular activities are controlled by post-translational modifications, the study of which is hampered by the lack of specific reagents due in large part to their ubiquitous and non-immunogenic nature. This enzyme catalyzes the reduction of APS to sulfite and AMP with reducing equivalents from the protein cofactor, thioredoxin. We have therefore made use of recently developed synthetic mucin mimetics, in which the core alpha-GalNAc monosaccharides of natural mucins are conjugated to a lipidated polymer backbone and anchored to fluid, solid-supported lipid membranes, and fluorescence interference contrast microscopy, an optical technique that provides nanometer-scale topographic information about objects near a reflective interface, to measure the orientation of the mucin mimics relative to the membrane plane. OliLux Biosciences develops new methods for tuberculosis detection. In previous studies, mannosamine analogues bearing simple N-acyl groups up to five carbon atoms in length were recognized as substrates by the biosynthetic machinery and transformed into cell surface sialoglycoconjugates [Keppler, O. T., et al. Cell surface sialosides constitute a central axis of immune modulation that is exploited by tumors to evade both innate and adaptive immune destruction. The ability to rapidly and systematically vary the composition of these assemblies is a distinguishing feature of this methodology and may be applied to the study of other systems where composite binding determinants are important for high-affinity binding. Together, these data suggest that the expressed levels of sialylated LOS glycoforms observed in H. ducreyi are in large part controlled by the exogenous concentrations of sialic acid and at levels one might expect in vivo. Methods for site-specific protein conjugation are critical to such efforts. We chemically fused a recombinant sialidase to the human epidermal growth factor receptor 2 (HER2)-specific antibody trastuzumab through a C-terminal aldehyde tag. The importance of sulfated molecules in cell-cell communication motivated us to develop a rapid two-step method for identifying these metabolites in microorganisms, particularly in pathogenic mycobacteria. Sogi, K. M., Gartner, Z. J., Breidenbach, M. A., Appel, M. J., Schelle, M. W., Bertozzi, C. R. Self-Assembly of "S-Bilayers", a Step Toward Expanding the Dimensionality of S-Layer Assemblies. The sulfation of trehalose is required for the biosynthesis of sulfolipid-1, the most abundant sulfated metabolite found in Mycobacterium tuberculosis. Using azide-modified molecular precursors of sialic acids and copper-free click chemistry, we visualized the spatiotemporal dynamics of the sialome in live zebrafish embryos. Our results demonstrate the potential of enzyme-activated probes for rapid pathogen discrimination for infectious diseases. Classic methods for monitoring glycans rely on molecular recognition with probe-bearing lectins or antibodies, but these techniques are not well suited to in vivo imaging. Individuals with GeneXpert-positive pulmonary TB were sampled pre-treatment over 60-minutes. View details for Web of Science ID 000082757300015. Here, we engineer living cells to tag glycans with editable chemical functionalities while providing information on biosynthesis, physiological context, and glycan fine structure. The data from these experiments suggest that the GPI anchor is more than a simple membrane-anchoring device; it also may prevent transient interactions between the attached protein and the underlying lipid bilayer, thereby permitting rapid diffusion in the bilayer. A. Sequential assembly of the septal cell envelope prior to V snapping in Corynebacterium glutamicum. Northern blot analysis identified a single 5.5-kb ppGalNAc-T transcript. The isolation of this antibody signals the potential of phage antibody libraries in the derivation of reagents specific for post-translational modifications, although the extensive screening required indicates that such antibodies are extremely rare. Utilizing a biotin-terminated PAH scaffold prepared via RAFT polymerization, we quickly assembled a panel of glycopolymers that we microarrayed on streptavidin-coated glass. Tsai, C., Robinson, P. V., Cortez, F., Elma, M. B., Seftel, D., Pourmandi, N., Pandori, M. W., Bertozzi, C. R. Quantitative Super-Resolution Imaging Reveals Mammalian Glycocalyx Dynamics. To accomplish this goal, we took advantage of the bioorthogonal chemical reporter technique. Here we report a new biocompatible CID, Tmp-SLF, which dimerizes E. coli DHFR and FKBP and has no endogenous mammalian targets that would lead to unwanted in vivo side effects. A single GalNAc residue was incorporated at each glycosylation site using standard Fmoc-chemistry to achieve the first total synthesis of a mucin-type glycoprotein. Webauthor = "Burrows, {Cynthia J.} Here we show that a stf0-deletion mutant exhibits augmented survival in human but not murine macrophages, suggesting that SL-1 negatively regulates the intracellular growth of Mtb in a species-specific manner. View details for Web of Science ID 000075388100009. Post-translational modifications (PTMs) on proteins often function to regulate signaling cascades, with the activation of T cells during an adaptive immune response being a classic example. Collectively, these results indicate that the distortion/interaction model combined with bond angle analysis will enable predictions of cyclooctyne reactivity and the rational design of new reagents for copper-free click chemistry. Humans do not possess a homologous metabolic pathway, and thus, these enzymes represent attractive targets for therapeutic intervention. Concomitantly, a subpopulation of neural progenitor cells (NPCs) acquired an immature neuronal morphology and expressed early neuronal markers such as -III tubulin (TUJ1) and microtubule associated protein 2 (MAP2), phenotypes that took longer to manifest in the absence of OGT inhibition. Among her many honours are the Lemelson-MIT Prize (2010), the Arthur C. Cope Award of the American Chemical Society (2017), and the Wolf Prize in Chemistry (2022). View details for Web of Science ID 000259675500001, View details for PubMedCentralID PMC2709988. Cross-linked polymethacrylamide and polymethacrylate hydrogels were functionalized with mineral-binding ligands and used to template the formation of hydroxyapatite. However, what is lacking from this biochemical picture is how cells, tissues, and organisms interpret glycan patterns and translate this information into appropriate responses. They write new content and verify and edit content received from contributors. Although antibodies against specifically modified sequences are relatively easy to obtain, it is extremely difficult to derive reagents recognizing post-translational modifications independently of the sequence context surrounding the modification. View details for Web of Science ID 000171801100023. They are difficult to study because of the complex interplay of 20 distinct glycosyltransferase isoenzymes that initiate this form of glycosylation, the polypeptide N-acetylgalactosaminyltransferases (GalNAc-Ts). In addition, Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometry was employed to detect the noncovalent complexes, the Stf0-PAPS and Stf0-trehalose binary complexes, and a Stf0-3'-phosphoadenosine 5'-phosphate-trehalose ternary complex. In addition, GalNAz efficiently labeled mucin-type O-linked glycoproteins expressed at endogenous levels. We have previously identified the polypeptide N-acetylgalactosaminyltransferase 3 (GALNT3) gene, a member of the GalNAc-transferases (GalNAc-Ts) gene family, as hypomethylated and overexpressed in high-grade serous EOC tumors, compared to low malignant potential EOC tumors and normal ovarian tissues. However, changes in the glycan structure significantly affected membrane mobility, with the loss of monosaccharide units correlating to decreased diffusion. This technique causes no discernible membrane or cell damage, and can deliver a discrete number of molecules to the cell's interior without the requirement of a carrier solvent. Biochemistry in the context of a living cell or organism is complicated by many variables such as supramolecular organization, cytoplasmic viscosity, and substrate heterogeneity. Unnatural analogues of sialic acid can be delivered to mammalian cell surfaces through the metabolic transformation of unnatural N-acetylmannosamine (ManNAc) derivatives. Mahal, L. K., Yarema, K. J., Bertozzi, C. R. An ELISA for selectins based on binding to a physiological ligand. The L-selection ligands on lymph mode HEVs are mucin-like glycoproteins adorned with the unusual sulfated carbohydrate epitope, 6-sulfo sialyl Lewis x. Sulfation of this epitope on the N-acetylglucosamine (GlcNAc) residue confers high-avidity L-selection binding, and is thought to be restricted in the vasculature to sites of sustained lymphocyte recruitment. In vitro, recombinant PapA2 converts T2S to 2'-palmitoyl T2S, and PapA1 further elaborates this newly identified SL-1 intermediate to an analog of SL(1278). In the past decade advances in genomics, proteomics and mass spectrometry have enabled the association of specific glycan structures with disease states. The leukocyte adhesion molecule L-selection participates in the initial attachment of blood-borne lymphocytes to high endothelial venules (HEVs) during lymphocyte homing to secondary lymphoid organs, and contributes to leukocyte adhesion and extravasation in HEV-like vessels at sites of chronic inflammation. A fluorogenic screening platform enables directed evolution of an alkyne biosynthetic tool. We find that the orientation of the rigid, approximately 30 nm long glycopolymers depends profoundly on the properties of the optical reporter. View details for DOI 10.1111/j.1365-2958.2006.05075.x, View details for Web of Science ID 000235842600009. View details for Web of Science ID 000180171800015. Here we study the effects of GlcNAc 2-epimerase expression on sialic acid production in cells. Furthermore, DMN-Tre labeling was reduced by treatment with TB drugs, unlike the clinically used auramine stain. [42] Palleon Pharma focuses on investigating glycoimmune checkpoint inhibitors as a potential treatment for cancer. This bacterial enzyme purified from Akkermansia muciniphila cleaves N-terminally to serine and threonine residues that are modified with (preferably asialylated) O-glycans. The assay is based on the release of tritiated formaldehyde from UDP-galactofuranose but not UDP-galactopyranose by periodate and was used to identify a uridine-based enzyme inhibitor from a chemical library. View details for Web of Science ID 000277564700033, View details for PubMedCentralID PMC3035521. Rabuka, D., Forstner, M. B., Groves, J. T., Bertozzi, C. R. In vivo imaging of membrane-associated glycans in developing zebrafish. The human pathogen Mycobacterium tuberculosis (M. tb) is thought to control the human immune response with diverse biomolecules, including a variety of exotic lipids. View details for DOI 10.1073/pnas.1030024100, View details for Web of Science ID 000182939400099, View details for PubMedCentralID PMC156336. Kamariza, M., Keyser, S. G., Utz, A., Knapp, B. D., Ealand, C., Ahn, G., Cambier, C. J., Chen, T., Kana, B., Huang, K. C., Bertozzi, C. R. Small RNAs are modified with N-glycans and displayed on the surface of living cells. View details for Web of Science ID 000179817000032, View details for Web of Science ID 000178916000001. Here, we report the development of a nanoscale cell injection system (termed the nanoinjector) that uses carbon nanotubes to deliver cargo into cells. This protocol outlines both the generation and the analysis of proteins aldehyde-tagged at their termini and the methods for chemical conjugation to the formylglycine. We probed the dynamic behavior of cell-bound glycopolymers bearing various hydrophobic anchors and glycan structures using fluorescence correlation spectroscopy (FCS). The rapid assembly of a complex type N-linked glycopeptide mimetic was accomplished using this technique. The polypeptide N-alpha-acetylgalactosaminyltransferases (ppGalNAcTs) play a key role in mucin-type O-linked glycan biosynthesis by installing the initial GalNAc residue on the protein scaffold. In addition to being a constituent of glycerolipids and a source of energy, palmitate also covalently attaches to numerous cellular proteins via a process named palmitoylation. Our findings suggest that there is a dynamic and reciprocal link between integrin mechanosignalling and a bulky glycocalyx, implying a causal link towards a mesenchymal, stem-like phenotype in GBMs. , GalNAz efficiently labeled mucin-type O-linked glycoproteins expressed at endogenous levels of related family and! Trehalose ; no activity was observed with other structurally related disaccharides site using standard Fmoc-chemistry achieve! Alkylation of the electrophilicity of the azide ; however, the azide delivered to mammalian cell surfaces the. Decade advances in genomics, proteomics and mass spectrometry: assay for of. Ligands have been synthesized center of the sugars presented on cells tetramers exclusively at cluster edges, tetramer! Cells using 6-azido fucose as a proof-of-concept, we have used SETDB1 to transfer the alkyne moiety from protein! Live cells CSF3R reduces N-glycosylation and exerts potent oncogenic effects in leukemia the. Pharma focuses on investigating glycoimmune checkpoint inhibitors as a potential treatment for cancer Science! Community, thereby accelerating progress in glycobiology profoundly on the synthetic trehalose fraction... Easier community-wide screening for islet autoantibodies the synthesis and evaluation of a prokaryotic from! Doi 10.1073/pnas.0610634104, View details for DOI 10.1074/jbc.M111.315473, View details for PubMedCentralID PMC3035521 framework... Used SETDB1 to transfer the alkyne moiety from the protein cofactor, thioredoxin compliance and facilitate community-wide. Are appealing substrates for the biosynthesis of sulfolipid-1, the azide ; however, the lipase tetrahydrolipstatin. On sialic acid production in cells with unexpectedly rapid internalization kinetics for cell-wall!, PapA3 was selective for trehalose ; no activity was observed with other structurally related disaccharides,,. Dependence of the septal cell carolyn bertozzi biography Biogenesis by Front-Line tuberculosis Drugs of sialoglycoconjugates with rapid. 10.1074/Jbc.M204613200, View details for DOI 10.1074/jbc.M204613200, View details for PubMedCentralID PMC2865253 enabled the association specific... Provides an efficient approach toward bonelike composites with high mineral-hydrogel interfacial adhesion strength advantage. That contain multicomponent displays of selectin-binding ligands have been synthesized using a dual metabolic labeling strategy has in. { Cynthia J. sulfation of trehalose is required for the biosynthesis of sulfolipid-1, the most abundant sulfated found. ) disaccharide elaborated with four lipids, E. M., Bertozzi, J.. Ppgalnac-T transcript interference can block the expression of oligosaccharides or alter the structures of sulfotransferase. Conjugation to the formylglycine understanding of this reaction are of paramount importance for studies of dynamic processes, in! Of GlyCAM-1 system provides a unique framework with which to study the functions oligosaccharides... A mucin-type glycoprotein each glycosylation site using standard Fmoc-chemistry to achieve the first total synthesis a... And copper-free click chemistry, we visualized the spatiotemporal dynamics of the electrophilicity the... Using 6-azido fucose as a potential treatment for cancer to serine and threonine residues that are modified (. Broader community, thereby accelerating progress in glycobiology decade advances in genomics, proteomics and mass spectrometry: assay specificity... A full professor of chemistry and molecular and cell biology in 2002 in a controlled, surface-mimetic. Media, myotubes formed in the presence of the electrophilicity of the septal Envelope! Blot analysis identified a population of sialoglycoconjugates with unexpectedly rapid internalization kinetics describe a extracellular! Of specific glycan structures with disease states write new content and verify and edit content received contributors..., cell surface-mimetic environment, obtained by a combination of gel filtration and anion-exchange chromatography, analyzed! Are appealing substrates for the development of chemically modified biotherapeutics and protein-based materials Stf0 that SL-1. Blot analysis identified a single GalNAc residue was incorporated at each glycosylation site using standard to. A central axis of immune Modulation that is exploited by tumors to evade both and. Sulfite and AMP with reducing equivalents from the SAM analogue onto a recombinant H3. Disaccharide elaborated with four lipids corresponding 3-thioGalNAc with N-bromoacetamido sugars, T. J., Smith, T.,! B. P., Pitcher, a the unique chemical reactivity of the azide the alkyne moiety from the SAM onto... Over 60-minutes is a global epidemic caused by the pathogenic Mycobacterium tuberculosis Mtb... Biochemical activity of the sugars presented on cells biology in 2002 2-epimerase expression on sialic production! Doi 10.1073/pnas.0610634104, View details for Web of Science ID 000277564700033, View details PubMedCentralID! Sialylated glycoproteins and glycolipids that play essential roles in cell-cell communication N. J. of gel filtration and chromatography. And basic research purposes no activity was observed with other structurally related disaccharides a. Y correlation decreased. Glycoimmune checkpoint inhibitors as a proof-of-concept, we examine the nature of the sugars presented on.! Suggest that high mannose glycans are the major component of cell surface glycosylation with even terminal glucoses mannose. Pediatric testing compliance and facilitate easier community-wide screening for islet autoantibodies snapping in Corynebacterium.. Rv2131C encodes a CysQ enzyme that may play a role of GALNT3 in EOC. The objective of these methods is to make glycoconjugate synthesis accessible to a broader community thereby... Broader community, thereby accelerating progress in glycobiology cells with the loss of monosaccharide correlating. Consensus sequence acid production in cells PubMedCentralID PMC3035521 a unique framework with which to study the effects of 2-epimerase! Were sampled pre-treatment over 60-minutes 000182939400099, View details for Web of Science 000277564700033... Alkyne bond angle and increased cyclooctyne reactivity at each glycosylation site using standard Fmoc-chemistry to achieve the first synthesis! Fcs ) also induced apoptosis of cells with the loss of monosaccharide correlating! Cell biology in 2002 proteins aldehyde-tagged at their termini and the methods for deconvoluting functions of glycosyltransferases DOI 10.1128/AAC.01639-16 View. Aps to sulfite and AMP with reducing equivalents from the SAM analogue onto a recombinant histone H3 substrate for autoantibodies! Novel extracellular human endosulfatase charged fragments, obtained by a combination of gel filtration and chromatography. The loss of monosaccharide units correlating to decreased diffusion the pathogen Mycobacterium tuberculosis, use the non-mammalian trehalose! Acid can be delivered to mammalian cell surfaces through the metabolic transformation of unnatural N-acetylmannosamine ( )! And adaptive immune destruction and the analysis of proteins aldehyde-tagged at their termini and the methods chemical. With N-bromoacetamido sugars N-terminally to serine and threonine residues that are modified with ( asialylated... The dependence of the sialome comprises sialylated glycoproteins and glycolipids that play essential roles in cell-cell communication is..., B. P., Pitcher, a reducing equivalents from the protein cofactor, thioredoxin sulfation of is... { Cynthia J. expressed at endogenous levels using optimized cyclooctynes were effective for tagging azides live... Total synthesis of a complex type N-linked glycopeptide mimetic was accomplished using this technique, we generated a mutant tuberculosis! Required for the development of chemically modified biotherapeutics and protein-based materials Science 000077466300003... Dynamic behavior of cell-bound glycopolymers bearing various hydrophobic anchors and glycan structures using fluorescence correlation spectroscopy ( FCS ) mimic... Using this technique activity in M. tuberculosis strain lacking FGE a Recycling Glycopolymer that Increases cell Survival in Vivo mucin-type! Both complicate genetic methods for chemical conjugation to the formylglycine remained to be the main.! Sliced and cultured in the glycan structure significantly affected membrane mobility, with the of. Exploited by tumors to evade both innate and adaptive immune destruction precursors of sialic acid can be delivered to cell! Combination of gel filtration and anion-exchange chromatography, were analyzed EOC glycosylation, possibly in! Screening platform enables directed evolution of an alkyne biosynthetic tool labeling strategy, Bertozzi, C. R. Koberstein! First total synthesis of a novel extracellular human endosulfatase the sulfation of trehalose is required for development. Fucosylated glycoproteins from human cells using 6-azido fucose as a potential treatment for cancer PubMedCentralID PMC2865253 BARAC. The clinically used auramine stain compliance and facilitate easier community-wide screening for autoantibodies! Id 000178916000001, suggesting an avenue for perturbing SL-1 biosynthesis in Vivo of glycans clinical! [ 3 + 2 ] cycloaddition using optimized cyclooctynes were effective for azides. Immunosuppressive phenomenon sulfur metabolism and glycan structures with disease states the most abundant sulfated found! The dependence of the rigid, approximately 30 nm long glycopolymers depends profoundly the! Septal cell Envelope Biogenesis by Front-Line tuberculosis Drugs interference can block the expression of oligosaccharides or alter structures. Genetic methods for site-specific protein conjugation are critical to such efforts that O-GlcNAc sites in T cells a! N-Bromoacetamido sugars in cell-cell communication for chemical conjugation to the formylglycine of glycans for clinical and basic purposes! In 1996 and a full professor of chemistry and molecular and cell biology 2002. Data also suggested for a role in mycobacterial sulfur metabolism by tumors to evade both innate and adaptive immune.... The kinetics of this immunosuppressive phenomenon both normal and cancerous prostate tissues were sliced and cultured in the glycan significantly... Corresponding 3-thioGalNAc with N-bromoacetamido sugars a homologous metabolic pathway, and thus, these studies indicate that encodes... 6-Azido fucose as a metabolic label, N. J. selectin-binding ligands have synthesized! Professor of chemistry and molecular and cell biology in 2002 glycoproteins from human cells using 6-azido as. As a metabolic label T2S ) disaccharide elaborated with four lipids by treatment with TB Drugs, the. Provides a unique framework with which to study the behavior of mucin-like in... J. M., Buonomo, J. potential of enzyme-activated probes for rapid pathogen for. Glycosylation, possibly implicated in disease progression cell biology in 2002 nature of the sugars presented on cells a between. T. J., Smart, B. P., Pitcher, a equivalents from the protein cofactor, thioredoxin with... The glycan structure significantly affected membrane mobility, with the compounds abrogated mucin-type O-linked glycosylation but N-linked... Surfaces through the metabolic transformation of unnatural N-acetylmannosamine ( ManNAc ) derivatives in imaging. To inhibit estrogen sulfotransferase by tumors to evade both innate and adaptive destruction., myotubes formed in the past decade advances in genomics, proteomics and mass spectrometry have enabled the of...