These studies indicate that copper-free crosslinking substrates uniquely offer a diagnostic probe for protein-protein interactions. One of the most abundant is sulfolipid-1 (SL-1), a tetraacyl-sulfotrehalose glycolipid. N-Carboxyanhydride Polymerization of Glycopolypeptides That Activate Antigen-Presenting Cells through Dectin-1 and Dectin-2. Low molecular weight and singly charged fragments, obtained by a combination of gel filtration and anion-exchange chromatography, were analyzed. Three piperidine-melamine glycodendrimers were synthesized via a chemoenzymatic route and attached to human growth hormone and the Fc region of human IgG. View details for DOI 10.1126/science.1155106, View details for Web of Science ID 000255454300046. Azido sugars are fed to cells and integrated by the glycan biosynthetic machinery into various glycoconjugates. We have demonstrated that the quadricyclane ligation is compatible with, and orthogonal to, strain-promoted azide-alkyne cycloaddition and oxime ligation chemistries by performing all three reactions in one pot on differentially functionalized protein substrates. Here, we report a method that combines the metabolic incorporation of a cyclooctyne-functionalized sialic acid derivative with a ligation reaction of a fluorogenic tetrazine, allowing for the imaging of sialylated glycoconjugates within live zebrafish embryos. We labeled LAP fusion proteins expressed in living mammalian cells with Cy3, Alexa Fluor 568 and biotin. B., Moore, K. L., Ellman, J. View details for Web of Science ID 000222420300014. Data from two independent probes, fluorophores conjugated directly to the polymer backbone and fluorescent proteins bound to the sugar groups, unexpectedly show that the mucin mimic molecules lie flat along the membrane. The approach was validated by labeling a recombinant glycoprotein that is known to possess O-linked glycans with GalNAz. We identified a concise disaccharide motif, GlcNAc beta 1-->6Gal alpha-R, that preserved both recognition and specificity determinants for the GlcNAc-6-0-sulfotransferase. These findings suggest that high mannose glycans are the major component of cell surface glycosylation with even terminal glucoses. These products represent a new class of engineered biosimilars bearing novel glycodendrimer structures. At elevated temperatures, however, the mutant was unable to proliferate even in the presence of trehalose. 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 10.1021/acs.jproteome.0c00218. Biography. Given the widespread occurrence of eukaryotic-like Ser/Thr protein kinases in bacteria, these proteins may play a broad role in bacterial osmosensing. Andres, L. M., Blong, I. W., Evans, A. C., Rumachik, N. G., Yamaguchi, T., Pham, N. D., Thompson, P., Kohler, J. J., Bertozzi, C. R. Bioorthogonal Labeling of Human Prostate Cancer Tissue Slice Cultures for Glycoproteomics. OliLux Biosciences develops new methods for tuberculosis detection. The oligonucleotides were introduced using metabolic carbohydrate engineering, which allowed reactive tailoring of the cell surface glycans for chemoselective bioconjugation. 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. GST-5 has recently been characterized as a novel GalNAc 6-O-sulfotransferase termed chondroitin 6-sulfotransferase-2 (Kitagawa, H., Fujita, M., Itio, N., and Sugahara K. (2000) J. Biol. After completing postdoctoral work at UCSF in the field of cellular immunology, she joined the UC Berkeley faculty in 1996. The modification modulates protein-protein interactions in the extracellular environment. View details for Web of Science ID 000177962200018. View details for DOI 10.1074/mcp.M600314-MCP200, View details for Web of Science ID 000242852000012. For her groundbreaking contributions to click chemistry and bioorthogonal chemistry, Bertozzi was awarded the 2022 Nobel Prize for Chemistry, which she shared with American chemist K. Barry Sharpless and Danish chemist Morten P. Meldal. Carolyn Bertozzi. A., Lo, A., Codelli, J. View details for DOI 10.1128/AEM.01537-10, View details for Web of Science ID 000284310500026, View details for PubMedCentralID PMC2988590, View details for DOI 10.1002/cbic.201000419, View details for Web of Science ID 000284050000005, View details for PubMedCentralID PMC3069858, View details for DOI 10.1002/cbic.201000115, View details for Web of Science ID 000284050000006, View details for PubMedCentralID PMC3069857. ScTyrY43G and MmPheT413G label overlapping but distinct proteomes in human cell lines, with broader proteome coverage upon their coexpression. Molecular level analysis of cell-surface phenomena could benefit from model systems comprising structurally defined components. The approach has applications in tissue-selective imaging of glycans for clinical and basic research purposes. While individual CD45 molecules moved randomly, they were displaced from the advancing phagocytic cup by an expanding diffusional barrier. Douglas, E. S., Chandra, R. A., Bertozzi, C. R., Mathies, R. A., Francis, M. B. Metabolic labeling of glycans with azido sugars and subsequent glycan-profiling and visualization via Staudinger ligation, Synthetic glycobiology: exploits in the Golgi compartment, Using phage display to select antibodies recognizing post-translational modifications independently of sequence context. The GPI anchor is a complex structure comprising a phosphoethanolamine linker, glycan core, and phospholipid tail. View details for DOI 10.1016/j.bmc.2016.05.050, View details for Web of Science ID 000385905800005, View details for PubMedCentralID PMC5052108. Glioblastoma multiforme (GBMs) are recurrent lethal brain tumours. The most potent examined, 1-68A, is a pH-dependent, two-step, covalent inhibitor of Escherichia coli LpxC that competes with UDP to bind the enzyme in the first step of inhibition. In particular, Bertozzi has advanced the understanding of cell surface oligosaccharides involved in cell recognition and inter-cellular communication. The effort was enabled by a new high-fidelity pattern-searching and glycopeptide validation algorithm termed IsoStamp v2.0, as well as by novel stable isotope probes. The same mineralization technique, when applied to a hydrogel that is less prone to surface hydrolysis, led to distinctly different mineralization patterns, in terms of both the extent of mineralization and the crystallinity of the apatite grown on the hydrogel surface. Luchansky, S. J., Hang, H. C., Saxon, E., Grunwell, J. R., Danielle, C. Y., Dube, D. H., Bertozzi, C. R. Drug targeting Mycobacterium tuberculosis cell wall synthesis: Development of a microtiter plate-based screen for UDP-galactopyranose mutase and identification of an inhibitor from a uridine-based library. Monomeric and multivalent oligosaccharides that bind to bacterial and viral receptors have been shown to abrogate infection by agents such as Helicobacter pilori, influenza virus and HIV. Advances in cellular imaging and in small molecule-controlled gene expression, signal transduction and cell surface modification are discussed in this review. View details for Web of Science ID A1996VV23700024, View details for Web of Science ID A1996VG14400059, View details for Web of Science ID A1996UA50800010, View details for Web of Science ID A1996TZ99700058, View details for Web of Science ID A1996UC44000017. The efficacy of these drugs is frequently undermined by acquired resistance. Despite decades of accumulated knowledge about proteins and their post-translational modifications (PTMs), numerous questions remain regarding their molecular composition and biological function. She coined the term "bioorthogonal A. Inverting family GH156 sialidases define an unusual catalytic motif for glycosidase action. Sialidase conjugation to trastuzumab enhanced ADCC against tumor cells expressing moderate levels of HER2, suggesting a therapeutic strategy for cancer patients with lower HER2 levels or inherent trastuzumab resistance. The Gal/GalNAc/GlcNAc-6-O-sulfotransferases (GSTs) are a recently discovered family of carbohydrate sulfotransferases that share significant sequence homology at the amino acid level and mediate a number of different biological processes such as leukocyte adhesion at sites of chronic inflammation. Their dense glycosylation is believed to confer structural rigidity as well as molecular extension beyond the glycocalyx, crucial to interaction with the cellular environment. View details for DOI 10.1073/pnas.0912081107, View details for Web of Science ID 000278549300009, View details for PubMedCentralID PMC2890823. However, their diverse structures, which are the key to their function, have hampered studies by biologists and chemists alike. A step in this direction will be the development of tools to identify metabolites that share common structural features. FGE has emerged as an enabling biotechnology tool due to the robust utility of the aldehyde product as a bioconjugation handle in recombinant proteins. Mycobacterium tuberculosis, the causative agent of tuberculosis, produces unique sulfated metabolites associated with virulence. Sialylated glycans are found at elevated levels in many types of cancer and have been implicated in disease progression. Here, we use methods that combine collision- and electron-based fragmentation to characterize the number of O-glycosites that are present in O-glycopeptides derived from the OpeRATOR digestion of four known O-glycoproteins. Welch Award in Chemistry (2022); The Dickson Prize in Medicine, University of Pittsburgh (2022); Dr H.P. View details for Web of Science ID 000268395000075, View details for PubMedCentralID PMC2716393. We synthesized a 1338 member library of uridine analogs directed to the epimerase by virtue of substrate mimicry. Exhaled air and bioaerosol volumes were estimated using continuous CO2 monitoring and airborne particle counting, respectively. More broadly, well-defined synthetic glycopolymers enable the integration of glycoconjugate structural and spatial diversity in a single microarray screening platform. Utilizing a biotin-terminated PAH scaffold prepared via RAFT polymerization, we quickly assembled a panel of glycopolymers that we microarrayed on streptavidin-coated glass. We used this strategy to construct a paracrine signaling network in isolated 3-dimensional microtissues. The assay was developed and validated in 7 distinct cohorts (n = 858) with the majority of the cohorts blinded prior to analysis. Here we study the effects of GlcNAc 2-epimerase expression on sialic acid production in cells. Further, we propose that the method reported here could find widespread use in investigating the functional consequences of O-GlcNAcylation. 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. Stanford, CA 94305Phone: (650) 723-2501Campus Map, UndergraduatesPh.D. This nucleotide sugar was readily accepted by fucosyltransferases and provided robust cell-surface labeling of fucosylated glycans, as determined by flow cytometry and confocal microscopy analysis. We found that N-azidoacetylgalactosamine (GalNAz) is converted by endogenous mammalian biosynthetic enzymes to UDP-GalNAz and then epimerized to UDP-N-azidoacetylglucosamine (GlcNAz). She is also an Investigator at the Howard Hughes Medical Institute (HHMI) and is the former Director of the Molecular Foundry. Using rational prediction analysis to recognize putative internal myristoylation sites in caspase-cleaved proteins combined with our nonradioactive chemical detection method, we identify 5 new post-translationally myristoylatable proteins (PKC epsilon, CD-IC2, Bap31, MST3, and the catalytic subunit of glutamate cysteine ligase). Our results demonstrate a key role of PIKfyve in the processing and presentation of antigens, which should be taken into consideration when targeting PIKfyve in autoimmune disease and cancer. The artificial receptor enhanced adenoviral vector binding and gene transfer to cells that are relatively resistant to adenovirus infection. A novel and efficient enzyme kinetics assay using electrospray ionization mass spectrometry was developed and applied to the bacterial carbohydrate sulfotransferase (NodST). Shes been out since the late 1980s, when being a lesbian could have jeopardized her career. View details for Web of Science ID 000237590400012. Agarwal, P., van der Weijden, J., Sletten, E. M., Rabuka, D., Bertozzi, C. R. Real-Time Noninvasive Imaging of Fatty Acid Uptake in Vivo. View details for Web of Science ID 000182331800019. Jacobs, C. L., Yarema, K. J., Mahal, L. K., Nauman, D. A., Charters, N. W., Bertozzi, C. R. Fmoc-based synthesis of peptide-(alpha)thioesters: Application to the total chemical synthesis of a glycoprotein by native chemical ligation. View details for Web of Science ID A1995TG85900001. How this multilayered cell envelope is assembled remains unclear. Sulfonucleotide reductases are a diverse family of enzymes that catalyze the first committed step of reductive sulfur assimilation. These cells were used as substrates to examine the effect of inhibiting PSA synthesis on the development of neurons derived from the chick dorsal root ganglion. The nematode Caenorhabditis elegans is an excellent model organism for studies of glycan dynamics, a goal that requires tools for imaging glycans in vivo. Detection of metabolites and post-translational modifications can be achieved using the azide as a bioorthogonal chemical reporter. Both the activity and specificity of the inducible enzymes were indistinguishable from their WT counterparts. and Shu Wang and Kim, {Hyun Jae} and Meyer, {Gerald J.} Surprisingly, Gal-1's effects on mammary patterning were independent of its glycan-binding ability and instead required localization within the nuclei of mammary epithelia. Methods for site-specific protein conjugation are critical to such efforts. Shieh, P., Siegrist, M. S., Cullen, A. J., Bertozzi, C. R. Glycocalyx engineering reveals a Siglec-based mechanism for NK cell immunoevasion. 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. Thus, this metabolic gene of an intracellular pathogen could have a secondary role in protection against the host immune response. This position statement originated from a working group meeting convened on April 15, 2015, by the NHLBI and incorporates follow-up contributions by the participants as well as other thought leaders subsequently consulted, who together represent research fields relevant to all branches of the NIH. That are relatively resistant to adenovirus infection achieved using the azide as a bioconjugation handle in recombinant.! Step in this direction will be the development of tools to identify metabolites that share structural. 3-Dimensional microtissues by a combination of gel filtration and anion-exchange chromatography, were analyzed displaced. Studies indicate that copper-free crosslinking substrates uniquely offer carolyn bertozzi biography diagnostic probe for protein-protein interactions of... 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