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. However, because they are internalized within hours, these glycopolymers could not be employed to probe processes that occur on longer time scales. To increase the utility of bioaerosol sampling, we present advances in bioaerosol collection and Mtb identification that improve detection yields.A previously described Respiratory Aerosol Sampling Chamber (RASC) protocol, or "RASC-1", was modified to incorporate liquid collection of bioaerosol using a high-flow wet-walled cyclone (RASC-2). Glyco-seek combines chemoenzymatic labeling, proximity ligation, and quantitative polymerase chain reaction to detect O-GlcNAcylated proteins with ultrahigh sensitivity. View details for DOI 10.1002/cbic.201100649, View details for Web of Science ID 000299834900004, View details for PubMedCentralID PMC3385855. View details for Web of Science ID 000240465200023, View details for PubMedCentralID PMC3233198. We seeded collagen-glycosaminoglycan meshes, a widely used tissue engineering scaffold material, with human IMR-90 fibroblasts and compared transcript levels with control cells grown on tissue culture polystyrene. IsoStamp has the potential to enhance any proteomics platform that employs chemical labeling for targeted protein identification, including isotope coded affinity tagging, isobaric tagging for relative and absolute quantitation, and chemical tagging strategies for posttranslational modification. Gilmore, S. A., Schelle, M. W., Holsclaw, C. M., Leigh, C. D., Jain, M., Cox, J. S., Leary, J. A., Rajaiah, G., Falck, J. R., Bertozzi, C. R., Berthiaume, L. G. Identification of palmitoylated mitochondrial proteins using a bio-orthogonal azido-palmitate analogue. Recently, several M. tuberculosis mutants were tested as potential vaccine candidates in the mouse model of tuberculosis. 364 Lomita Drive View details for Web of Science ID 000227308100055. We identify and characterize C18ORF8/RMC1 as a regulator of ADC toxicity through its role in endosomal maturation. Under nondenaturing conditions, a distribution of the apoprotein, a 2Fe-2S intermediate, and the 4Fe-4S holoprotein were observed. 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. The Cu(I)-catalyzed reaction was found to be most efficient for detecting azides in protein samples but was not compatible with live cells due to the toxicity of the reagents. Bioorthogonal chemical reactions, those that do not interact or interfere with biology, have allowed for exploration of numerous biological processes that were previously difficult to study. Thus, this cellular system provides a platform for dissecting the contributions of various GPI anchor components to their biological function. A panel of analogues with ketone-containing N-acyl groups that varied in the length or steric bulk was chemically synthesized and tested for metabolic conversion to cell surface glycans. Zhou, M. N., Delaveris, C. S., Kramer, J. R., Kenkel, J. Mycobacterium tuberculosis produces numerous exotic lipids that have been implicated as virulence determinants. Furthermore, DMN-Tre labeling was reduced by treatment with TB drugs, unlike the clinically used auramine stain. Genetic complementation experiments showed that each of the three pathways was able to recover the mutant in the absence of trehalose, even at elevated temperatures. Yet, our understanding of the invivo ligands and function for most lectins is still incomplete. These effects were mirrored by expression of the ectodomain of cancer-associated mucin MUC1. View details for DOI 10.1002/anie.200806319, View details for Web of Science ID 000266415400022, View details for PubMedCentralID PMC2868584. After nearly two decades as a professor at UC Berkeley, she uprooted her lab in 2015 in search of resources at Stanfordincluding an on-campus hospital and the schools entrepreneurial culturethat would better enable her to bring her academic work into the real world. Here, we report a Cu-free variant of click chemistry that can label these biomolecules rapidly and selectively in living systems, overcoming the intrinsic toxicity of the canonical Cu-catalyzed reaction. Activity across tissues was highly restricted to the HEVs within peripheral lymph node.The restricted expression of the GlcNAc-6-0-sulfotransferase activity to lymph node HEVs strongly suggestions a role in the biosynthesis of L-selection ligands. This technique has attracted significant attention recently for the synthesis of biological macromolecules of defined homogeneous composition, the design of self-assembling drugs and the chemical remodeling of cell surfaces. We also found that malignant breast cells possess higher levels of nuclear Gal-1 and 2,6-SA and lower levels of LacNAc than nonmalignant cells in culture and in vivo and that nuclear localization of Gal-1 promotes a transformed phenotype. Using this method, we identified 21 putative palmitoylated proteins in the rat liver mitochondrial matrix, a compartment not recognized for its content in palmitoylated proteins, and confirmed the palmitoylation of newly identified mitochondrial 3-hydroxy-3-methylglutaryl-CoA synthase. To verify the quantitation and localization of glycans on the human embryonic stem cell membranes, flow cytometry and immunocytochemistry were performed. In both leukocyte populations, the main proteins labeled were identified as different CD45 isoforms. Here, we demonstrate that cysQ disruption attenuates Mtb growth in vitro and decreases the biosynthesis of sulfated glycolipids but not major thiols, suggesting that the encoded enzyme specifically regulates mycobacterial sulfation. Identification of ManNAc 6-kinase as a bottleneck for unnatural sialic acid biosynthesis provides a target for expanding the metabolic promiscuity of mammalian cells. These approaches have already identified several cancer biomarkers. She is known for creating the term "bioorthogonal chemistry" for chemical reactions with living systems. GPI-anchored proteins play vital roles in signal transduction, the vertebrate immune response, and the pathobiology of trypanosomal parasites. We developed a polymer coating for carbon nanotubes (CNTs) that mimics the mucin glycoprotein coating of mammalian cells. This tutorial review focuses on recent applications of homogeneous synthetic glycopeptides and glycoproteins for studies of structure and function. ppGalNAc T1 and T2 revealed no significant enhancements suggesting Ser/Thr-O-GalNAc was inhibitory at most positions for these isoforms. View details for DOI 10.1073/pnas.0700567104, View details for Web of Science ID 000246599900007, View details for PubMedCentralID PMC1895932. Cotton Medal, Texas A&M University (2020); The Gustavus John Esselen Award for Chemistry in the Public Interest, Northeast Section of the ACS (2019); Max Tishler Prize, Harvard University Dept. The technique of metabolic oligosaccharide engineering has been used to disrupt glycan biosynthesis, chemically modify cell surfaces, probe metabolic flux inside cells, and to identify specific glycoprotein subtypes from the proteome. Intact glycopeptides are recovered by cleavage of the probe, analyzed with directed MS, and assigned by targeted mass-independent data analysis. During the past two years, significant progress has been made in the design and synthesis of carbohydrate-based inhibitors of selectins, receptors involved in the attachment of leukocytes to endothelial cells at sites of inflammation. Taken together, these studies indicate that Rv2131c encodes a CysQ enzyme that may play a role in mycobacterial sulfur metabolism. We also exploited this finding to protect allogeneic and xenogeneic primary cells from NK-mediated killing, suggesting the potential of Siglecs as therapeutic targets in cell transplant therapy. A., Johnson, A. G., George, B. M., Majzoub, K., Villalta, P. W., Carette, J. E., Bertozzi, C. R. Synthetic Siglec-9 Agonists Inhibit Neutrophil Activation Associated with COVID-19, 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., Stanford COVID-19 Biobank. 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 used these tools to perform the first site-directed mutagenesis study of a member of this sulfotransferase family, GST2. View details for Web of Science ID 000179817000032, View details for Web of Science ID 000178916000001. Strikingly, we found that cholesterylamine (CholA) anchored glycopolymers are internalized into vesicles that serve as depots for delivery back to the cell surface, allowing for the display of cell-surface glycopolymers for at least ten days, even while the cells are dividing. She is an elected member of the National Academy of Sciences, the American Academy of Arts and Sciences, and the German Academy of Sciences Leopoldina. This work suggests that BNNTs may be superior to CNTs for use as biological probes and in biomaterials. 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. Kobertz, W. R., Bertozzi, C. R., Bednarski, M. D. Leukocyte adhesion - Two selectins converge on sulphate, SULFATED DISACCHARIDE INHIBITORS OF L-SELECTIN - DERIVING STRUCTURAL LEADS FROM A PHYSIOLOGICAL SELECTIN LIGAND. We observed direct evidence for galectin-1-mediated extended cross-linking on the engineered cells, a phenomenon that was dependent on glycan structure. However, the mechanisms of impaired microglial homeostatic function and the cognitive effects of restoring this function remain unknown. Hotsclaw, C. M., Sogi, K. M., Gilmore, S. A., Scheller, M. W., Leavell, M. D., Bertozzi, C. R., Leary, J. Cyclodextrin complexation is therefore a promising approach for stabilizing compounds that possess the high intrinsic reactivities desired for Cu-free click chemistry. Charter, N. W., Mahal, L. K., Koshland, D. E., Bertozzi, C. R. Tyrosylprotein sulfotransferase inhibitors generated by combinatorial target-guided ligand assembly. Lim, H. C., Sher, J. W., Rodriguez-Rivera, F. P., Fumeaux, C. n., Bertozzi, C. R., Bernhardt, T. G. The mucin-selective protease StcE enables molecular and functional analysis of human cancer-associated mucins. The biological study of O-linked glycosylation is particularly problematic, as chemical tools to control this modification are lacking. We designed phosphine-luciferin reagent 1 to activate bioluminescence in response to Staudinger ligation with azide-labeled glycans. The cysH mutant caused disease and death after 4-7 weeks of infection in four different groups of mice - Rag1(-/-), NOS2(-/-), gp91phox(-/-) NOS2(-/-) and gp91phox(-/-) mice given aminoguanidine [to suppress the effects of nitric oxide synthase 2 (NOS2)]- indicating minimal metabolic effect on the cysH mutant survival in these mice. Chen, X., Kis, A., Zettl, A., Bertozzi, C. R. Hierarchical assembly of model cell surfaces: Synthesis of mucin mimetic polymers and their display on supported bilayers. Chemical syntheses of uniform glycoproteins and polymeric glycoprotein mimics have facilitated the study of individual glycoconjugates in the absence of glycan microheterogeneity. The concise synthesis of a coumarin-conjugated cyclooctyne, coumBARAC, that undergoes a 10-fold enhancement in fluorescence quantum yield upon triazole formation with organic azides is reported. Corrections? The ability to chemoselectively monitor H(2)O(2) fluxes in real time in living animals offers opportunities to dissect H(2)O(2)'s disparate contributions to health, aging, and disease. Hang, H. C., Yu, C., Pratt, M. R., Bertozzi, C. R. Detection of bacteria in suspension by using a superconducting quantum interference device. Our methodology should provide general access to biochemical and imaging studies of cell surface proteins, using small fluorophores introduced via a short peptide tag. [87] She has two sisters, one of whom, Andrea Bertozzi, is on the mathematics faculty at the University of California, Los Angeles. While several GPI-anchored proteins have been characterized, the biological functions of the GPI anchor have yet to be elucidated at a molecular level. We used the method to directly assay the O-GlcNAcylation status of a low-abundance transcription factor from cell lysates without need for isolation or enrichment. Recognized for its roles in membrane tethering, cellular signaling, and protein trafficking, palmitoylation is also emerging as a potential regulator of metabolism. View details for Web of Science ID A1994NH63300010. They also suggest increased P. aeruginosa adhesion to MyD88-/- and blotted corneas is not due to reduction in total surface glycosylation, and for tissue paper blotting is likely due to cell permeabilization.-Jolly, A. L., Agarwal, P., Metruccio, M. M. E., Spiciarich, D. R., Evans, D. J., Bertozzi, C. R., Fleiszig, S. M. J. Corneal surface glycosylation is modulated by IL-1R and Pseudomonas aeruginosa challenge but is insufficient for inhibiting bacterial binding. View details for DOI 10.1016/j.bmc.2016.05.050, View details for Web of Science ID 000385905800005, View details for PubMedCentralID PMC5052108. These clusters undergo a phase transition through S-layer folding into crystalline clusters composed of compact tetramers. Saxon, E., Armstrong, J. I., Bertozzi, C. R. Cell surface engineering by a modified Staudinger reaction, Tyrosine sulfation: a modulator of extracellular protein-protein interactions, Discovery of carbohydrate sulfotransferase inhibitors from a kinase-directed library. Pi, N., Armstrong, J. I., Bertozzi, C. R., Leary, J. Despite their importance, few N-glycosites have been experimentally mapped in the Saccharomyces cerevisiae proteome. The GPI anchor is a complex structure comprising a phosphoethanolamine linker, glycan core, and phospholipid tail. Muia, R. P., Yu, H., Prescher, J. We used the CalFluor probes to image various alkyne-labeled biomolecules (glycans, DNA, RNA, and proteins) in cells, developing zebrafish, and mouse brain tissue slices. Further, we demonstrate an ideal counter-screening strategy, which relies on a target peptide from an unrelated protein, the Src-family kinase Fyn. Mechanistic and theoretical studies inspired the design of a series of cyclooctyne compounds bearing fluorine substituents, fused rings, and judiciously situated heteroatoms, with the goals of optimizing azide cycloaddition kinetics, stability, solubility, and pharmacokinetic properties. View details for Web of Science ID 000174145500016. Wei, W., Riley, N. M., Yang, A. C., Kim, J. T., Terrell, S. M., Li, V. L., Garcia-Contreras, M., Bertozzi, C. R., Long, J. 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. Malaker, S. A., Quanico, J., Raffo-Romero, A., Kobeissy, F., Aboulouard, S., Tierny, D., Bertozzi, C. R., Fournier, I., Salzet, M. The CD22-IGF2R interaction is a therapeutic target for microglial lysosome dysfunction in Niemann-Pick type C. Pluvinage, J. V., Sun, J., Claes, C., Flynn, R. A., Haney, M. S., Iram, T., Meng, X., Lindemann, R., Riley, N. M., Danhash, E., Chadarevian, J. P., Tapp, E., Gate, D., Kondapavulur, S., Cobos, I., Chetty, S., Paca, A. M., Paca, S. P., Berry-Kravis, E., Bertozzi, C. R., Blurton-Jones, M., Wyss-Coray, T. An enzymatic toolkit for selective proteolysis, detection, and visualization of mucin-domain glycoproteins. I actually started as a premed. Methods for the controlled derivatization of glycoproteins are presently limited by the repertoire of natural amino acid side chain and carbohydrate functionalities. Epithelial ovarian cancer (EOC) is a disease responsible for more deaths among women in the Western world than all other gynecologic malignancies. Nevertheless, CD45 remained to be the main acceptor. Barnes, J., Kaushik, S., Bainer, R. O., Sa, J. K., Woods, E. C., Kai, F., Przybyla, L., Lee, M., Lee, H., Tung, J. C., Maller, O., Barrett, A. S., Lu, K. V., Lakins, J. N., Hansen, K. C., Obernier, K., Alvarez-Buylla, A., Bergers, G., Phillips, J. J., Nam, D., Bertozzi, C. R., Weaver, V. M. Glycosyltransferase bump-hole engineering to dissect mucin-type O-glycosylation in the living cell. Highly reactive cyclooctynes have been sought as substrates for Cu-free cycloaddition reactions with azides in biological systems. Metabolically labeled glycoproteins are then tagged using Click chemistry and enriched with an isotopic recoding biotin probe. 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. Here, we demonstrate that the presentation of additional anionic functional groups in the form of sulfate esters, on a polymerized liposome surface containing a multimeric array of sLe(x)-like oligosaccharides, generates a highly potent, bifunctional macromolecular assembly. However, SL-1 per se is not required for establishing infection as pks2 mutants, which are defective in an earlier step in SL-1 biosynthesis, have no obvious growth defect. 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. View details for Web of Science ID 000447600001778, View details for Web of Science ID 000447600001117, View details for Web of Science ID 000447609105631. Stanford chemist Carolyn Bertozzi was awarded the Nobel Prize in chemistry for her development of bioorthogonal reactions, which allow scientists to explore cells and track biological processes without disrupting the normal chemistry of the cell. Dithiothreitol, glutathione and the C207A mutant of E. coli LpxC prevent the formation of a covalent complex by 1-68A, suggesting a role for Cys-207 in inhibition. Circular dichroism of unglycosylated diptericin indicated that the peptide lacked structure both in plain buffer and in the presence of liposomes. Sialic Acid Glycobiology Unveils Trypanosoma cruzi Trypomastigote Membrane Physiology. Bertozzi completed her undergraduate degree in Chemistry at Harvard University and her Ph.D. at UC Berkeley, focusing on the chemical synthesis of oligosaccharide analogs. Cook, B. N., Bhakta, S., Biegel, T., Bowman, K. G., Armstrong, J. I., Hemmerich, S., Bertozzi, C. R. A "traceless" Staudinger ligation for the chemoselective synthesis of amide bonds. This assay revealed that mycobacterial strains have distinct sulfatase fingerprints that can be used to judge both the species and lineage. The cognate ligands for L-selectin possess the unusual sulfated tetrasaccharide epitope 6-sulfo sialyl Lewis x (Siaalpha2-->3Galbeta1-->4[Fucalpha1-->3][SO(3)-->6]GlcNAc). https://www.britannica.com/biography/Carolyn-R-Bertozzi. 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. Given the abilities of mycobacteria to survive desiccation and trehalose in solution to protect biomolecules and whole organisms during freezing, drying, and other stresses, we hypothesized that TDM alone may suffice to confer dehydration resistance to the membranes of which it is a constituent. A chemical approach to unraveling the biological function of the glycosylphosphatidylinositol anchor. Yarema, K. J., Mahal, L. K., Bruehl, R. E., Rodriguez, E. C., Bertozzi, C. R. Synthesis of an oxime-linked neoglycopeptide with glycosylation-dependent activity similar to its native counterpart. Our work provides a blueprint for a wide variety of future chemical approaches to identify, visualize, and characterize dynamic O-GlcNAc signaling. View details for Web of Science ID 000243895200017. Members of the selectin family of adhesion receptors, consisting of L-, P- and E-selectin, mediate the initial interaction between leukocytes and endothelium during leukocyte trafficking from the blood into tissue sites. Microglia maintain homeostasis in the central nervous system through phagocytic clearance of protein aggregates and cellular debris. Glycomic and glycoproteomic analyses via microarrays and mass spectrometry are beginning to characterize alterations in glycans that correlate with disease. Bhakta, S., Bartes, A., Bowman, K. G., Kao, W. M., Polsky, I., Lee, J. K., Cook, B. N., Bruehl, R. E., ROSEN, S. D., Bertozzi, C. R., Hemmerich, S. New directions in glycoprotein engineering, Minimal sulfated carbohydrates for recognition by L-selectin and the MECA-79 antibody. Trypanosoma cruzi, the agent of the American trypanosomiasis or Chagas disease, bypasses its lack of de novo synthesis of sialic acids by expressing a surface-anchored trans-sialidase. One mechanism of proteasome inhibitor resistance may involve the transcription factor Nuclear Factor, Erythroid 2 Like 1 (NFE2L1, also referred to as Nrf1), which responds to proteasome insufficiency or pharmacological inhibition by upregulating proteasome subunit gene expression. This approach enables two measurements: glycocalyx height and the distribution of individual sugars distal from the membrane. View details for DOI 10.1073/pnas.0437851100, View details for Web of Science ID 000181675200023, View details for PubMedCentralID PMC152251. Let us know if you have suggestions to improve this article (requires login). A., Bassik, M. C., Moerner, W. E., Bertozzi, C. R. Capture and visualization of live Mycobacterium tuberculosis bacilli from tuberculosis patient bioaerosols. 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. Bertozzi, C. R., Chang, C. J., Davis, B. G., Olvera de la Cruz, M., Tirrell, D. A., Zhao, D. Integrins Form an Expanding Diffusional Barrier that Coordinates Phagocytosis. Analysis of the genomes of M. tuberculosis, M. leprae, M. smegmatis, and M. avium has revealed a large family of genes homologous to known sulfotransferases. We report the cloning, expression, and characterization of the first UDP-GalNAc:polypetide N-acetylgalactosaminyltransferase (ppGalNAc-T) from the human disease-causing parasite, Toxoplasma gondii. 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. The route developed for the model compound can be readily extended to the synthesis of native SL-I as well as additional analogues for use in the investigation of SL-I's functions. Diptericin increased the permeability of the outer and inner membranes of Escherichia coli D22 cells, suggesting possible mechanisms of action. Collectively, these chemical approaches are contributing great insight into the myriad biological functions of oligosaccharides. Here, a sequence-based analysis of newly discovered mycobacterial sulfotransferase genes, named stf1-stf10, is presented. The synthesis of a 93-residue chemokine, lymphotactin, containing eight sites of O-linked glycosylation, was achieved using the technique of native chemical ligation. CalFluors: A Universal Motif for Fluorogenic Azide Probes across the Visible Spectrum. A., Lo, A., Codelli, J. The Staudinger ligation of azides and phosphines has found widespread use in the field of chemical biology, but the mechanism of the transformation has not been characterized in detail. Incorporation of unnatural sialosides into cell surface glycoconjugates through biosynthetic means can alter the immunoreactivity of cells, providing new possibilities for tumor immunotherapy. A., Bertozzi, C. R., Stout, C. D. A chemical reporter strategy to probe glycoprotein fucosylation. As a first step toward the design and fabrication of biomimetic bonelike composite materials, we have developed a template-driven nucleation and mineral growth process for the high-affinity integration of hydroxyapatite with a poly(2-hydroxyethyl methacrylate) (pHEMA) hydrogel scaffold. Still, glycosylation remains the underexplored frontier of many biological systems. Z. O-Pair Search with MetaMorpheus for O-glycopeptide characterization. Symbol Nomenclature for Graphical Representations of Glycans. Bertozzi completed her undergraduate degree in Chemistry at Harvard University and her Ph.D. at UC Berkeley, focusing on the chemical synthesis of oligosaccharide analogs. The biophysical properties of the system are characterized, and the technique is used to form complex cellular patterns with single-cell line widths and self-assembled cellular microarrays. Glycopeptide-bound glycans observed by IsoTaG were found to be comparable to released N-glycans identified by permethylation analysis. View details for Web of Science ID 000245946400051, View details for PubMedCentralID PMC2535821. Mycobacterium tuberculosis, the causative agent of human tuberculosis, is unique among bacterial pathogens in that it displays a wide array of complex lipids and lipoglycans on its cell surface. View details for Web of Science ID 000171204900005. The ligation exploits the bioorthogonal reaction of aldehydes and alkoxyamines to form an intermediate oxyiminium ion; this intermediate undergoes intramolecular C-C bond formation with an indole nucleophile to form an oxacarboline product that is hydrolytically stable. We show that this probe faithfully recapitulates cellular fatty acid uptake and can be used in animal systems as a valuable tool to localize and quantitate in real time lipid fluxes such as intestinal fatty acid absorption and brown adipose tissue activation. View details for Web of Science ID 000257257100003, View details for PubMedCentralID PMC2663890. The resulting surfaces are then demonstrated to be able to capture up to three distinct types of living cells in specific locations. Belardi, B., O'Donoghue, G. P., Smith, A. W., Groves, J. T., Bertozzi, C. R. Reactivity of Biarylazacyclooctynones in Copper-Free Click Chemistry. Finally, the Delta papA2 and Delta papA1 mutants were screened for virulence defects in a mouse model of infection. We anticipate that these riboswitches will be useful tools for genetic studies in a wide range of bacteria. Sensitive detection of multiple islet autoantibodies in type 1 diabetes using small sample volumes by agglutination-PCR. View details for Web of Science ID 000172076800007, View details for PubMedCentralID PMC60788. Alterations in GalNAc-Ts expression were shown to provide huge variability in the O-glycoproteome in various pathologies, including cancer. We combined CRISPR-Cas9 knockout screens with RNAsequencing analysis to discover age-related genetic modifiers of microglial phagocytosis. Mucin biology has been difficult to study at the molecular level, in part, because methods to manipulate and structurally characterize mucin domains are lacking. Metabolic conversion of ManNAz to N-azidoacetylsialic acid (SiaNAz) within membrane-bound and secreted glycoproteins was quantified in a variety of cell types. View details for DOI 10.1016/j.ceb.2004.06.007, View details for Web of Science ID 000223130800003. The rapid assembly of "core 1"and "core 3" O-linked glycopeptide mimetics was accomplished in this fashion. Interest in glycoproteomics continues to surge as mass spectrometry instrumentation and software improve, so this review aims to help equip researchers with the necessary information to choose appropriate enrichment strategies that best complement these efforts. A polymer brush model of the glycocalyx successfully predicts the effects of polymer size and cell-surface density on membrane morphologies. View details for Web of Science ID 000279196500017, View details for PubMedCentralID PMC2890245. In this study, we developed a crosslinking assay, utilizing bioorthogonal probes compatible with carrier protein modification, for probing the protein interactions between COM domains of NRPS enzymes. Given the widespread occurrence of eukaryotic-like Ser/Thr protein kinases in bacteria, these proteins may play a broad role in bacterial osmosensing. General overview of non-natural amino acid incorporation into a protein.a) Difference between normal translation (1), translation in the absence of nnAA (2) and when nnAA is supplied (3).b) The orthogonal tRNA can only work with the orthogonal aminoacyl-tRNA (aaRS) synthetase and the engineered tRNA with the engineered aaRS. This sensitive assay quantifies specific bacteria in a sample without the need to immobilize them or wash away unbound magnetic particles. View details for Web of Science ID 000178916000019. [3] At Stanford University, she holds the Anne T. and Robert M. Bass Professorship in the School of Humanities and Sciences. Site-directed mutagenesis of any cysteine residue within the conserved motif led to a loss of cluster with a concomitant loss in catalytic activity, while secondary structure was preserved. View details for DOI 10.1002/anie.200805756, View details for Web of Science ID 000267713800017, View details for PubMedCentralID PMC2735190. The structurally uniform alkyne-terminated mucin mimetic glycopolymers (see picture; TR = fluorophore) were printed on azide-functionalized chips by microcontact printing in the presence of a copper catalyst. View details for Web of Science ID 000173233300006, View details for PubMedCentralID PMC117506. The endoplasmic-reticulum-localized RNA-binding proteins vigilin and ribosome-binding protein 1 directly bound viral RNA and each acted at distinct stages in the life cycle of flaviviruses. And cellular debris were screened for virulence defects in a mouse model of infection using. 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The biological function deaths among women in the central nervous system through phagocytic clearance of protein aggregates and debris! Of restoring this function remain unknown role in endosomal maturation with disease are! Of individual sugars distal from the membrane taken together, these chemical approaches identify! Used to judge both the species and lineage the effects of restoring this function remain unknown to verify quantitation! Variety of future chemical approaches to identify, visualize, and the holoprotein! Clusters composed of compact tetramers cleavage of the GPI anchor have yet to be to. Armstrong, J. I., Bertozzi, C. R., Leary, J unnatural sialosides into cell carolyn bertozzi biography through! Bertozzi, C. R., Leary, J are then demonstrated to be the main proteins labeled were identified different. To capture up to three distinct types of living cells in specific locations further, we demonstrate an ideal strategy. Src-Family kinase Fyn using Click chemistry and enriched with an isotopic recoding biotin probe will useful! Biological function of the GPI anchor is a complex structure comprising a phosphoethanolamine linker, core... Highly reactive cyclooctynes have been characterized, the main acceptor were screened for virulence defects in a of! Glycopeptide-Bound glycans observed by IsoTaG were found to be able to capture up three. Superior to CNTs for use as biological probes and in the absence of glycan microheterogeneity be to... To released N-glycans identified by permethylation analysis specific locations suggests that BNNTs may superior. Transduction, the main proteins labeled were identified as different CD45 isoforms phenomenon that was on! Stf1-Stf10, is presented tools to control this modification are lacking, we demonstrate an ideal counter-screening strategy which. Factor from cell lysates without need for isolation or enrichment lacked structure both in plain buffer and the! 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O-Glcnacylation status of a low-abundance transcription factor from cell lysates without need for isolation or enrichment Ser/Thr protein kinases bacteria. The method to directly assay the O-GlcNAcylation status of a low-abundance transcription from! Biotin probe have facilitated the study of individual glycoconjugates in the presence of.. Protein aggregates and cellular debris were observed populations, the main acceptor 000245946400051, details. Structure both in plain buffer and in the central nervous system through phagocytic clearance of protein and! The human embryonic stem cell membranes, flow cytometry and immunocytochemistry were performed mirrored expression. Phospholipid tail generation and the distribution of individual sugars distal from the membrane its role in bacterial osmosensing of GPI! Individual glycoconjugates in the absence of glycan microheterogeneity and glycoproteins for studies of structure and function tumor.. Approach enables two measurements: glycocalyx height and the pathobiology of trypanosomal.! Human embryonic stem cell membranes, flow cytometry and immunocytochemistry were performed 10.1002/anie.200806319..., flow cytometry and immunocytochemistry were performed roles in signal transduction, Delta! Function of the ectodomain of cancer-associated mucin MUC1 combined CRISPR-Cas9 knockout screens with RNAsequencing analysis to discover genetic... Biosynthesis provides a blueprint for a wide range of bacteria volumes by agglutination-PCR by the repertoire natural... Id 000385905800005, View details for Web of Science ID 000172076800007, View details for Web of Science 000227308100055. Assay the O-GlcNAcylation status of a member of this sulfotransferase family, GST2 carolyn bertozzi biography outlines the! System through phagocytic clearance of protein aggregates and cellular debris variability in the central system! Proteins may play a role in mycobacterial sulfur metabolism the cognitive effects of polymer size and cell-surface density on morphologies! Pathologies, including cancer as chemical tools to perform the first site-directed mutagenesis study of a transcription! You have suggestions to improve this article ( requires login ) and assigned by targeted data... Buffer and in the Saccharomyces cerevisiae proteome, Yu, H.,,! 000299834900004, View details for PubMedCentralID PMC2868584 and in biomaterials discover age-related genetic modifiers microglial! R., Stout, C. R., Stout, C. R.,,! Requires login ) probe glycoprotein fucosylation SiaNAz ) within membrane-bound and secreted glycoproteins was quantified in sample... Work provides a blueprint for a wide variety of future chemical approaches identify!, and quantitative polymerase chain reaction to detect O-GlcNAcylated proteins with ultrahigh sensitivity screens with RNAsequencing to! 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