Pluvinage, J. V., Haney, M. S., Smith, B. The installation of sulfate groups on the carbohydrate residues of glycoproteins, glycolipids, and glycosaminoglycans is a critical posttranslational modification that occurs in all higher eukaryotes. Bifunctional, fluorinated cyclooctynes were used for the in situ "click" crosslinking of azide-terminated photodegradable star polymers, yielding photodegradable polymeric model networks with well-defined structures and tunable gelation times. Jolly, A. L., Agarwal, P., Metruccio, M. M., Spiciarich, D. R., Evans, D. J., Bertozzi, C. R., Fleiszig, S. M. Visualization of mycobacterial membrane dynamics in live cells. In the measurement, we add 50-nm-diameter superparamagnetic magnetite particles, coated with antibodies, to an aqueous sample containing L. monocytogenes. a nanoscience research center at Lawrence Berkeley National Laboratory. View details for DOI 10.1021/acs.jproteome.0c00218. We engineered mammalian cells to produce diazirine-modified O-GlcNAc by expressing a mutant form of UDP-GlcNAc pyrophosphorylase and subsequently culturing these cells with a cell-permeable, diazirine-modified form of GlcNAc-1-phosphate. Cell surface trehalose mycolates are important modulators of mycobacterial pathogenesis and host immune response. A comparative study of bioorthogonal reactions with azides. We identified prominent localization of mucins in the pharynx of all four larval stages, in the adult hermaphrodite pharynx, vulva and anus, and in the tail of the adult male. In this approach, metabolic labeling substrates containing bioorthogonal functional groups are provided to cells for incorporation into biopolymers by endogenous biosynthetic machinery. Our results show that, like palmitate, incorporation of azido-palmitate occurred on mitochondrial proteins via thioester bonds at sites that could be competed out by palmitoyl-CoA. View details for Web of Science ID 000169081700029. Yu, S., Boyce, M., Wands, A. M., Bond, M. R., Bertozzi, C. R., Kohler, J. J. Elucidation and Chemical Modulation of Sulfolipid-1 Biosynthesis in Mycobacterium tuberculosis. Lantos, A. Mougous, J. D., Green, R. E., Williams, S. J., Brenner, S. E., Bertozzi, C. R. Recent advances in the chemical synthesis of mucin-like glycoproteins, Conjugation of DNA to silanized colloidal semiconductor nanocrystalline quantum dots. [6] Her lab's development of nanotechnologies which probe biological systems lead to the development of a fast point-of-care tuberculosis test in 2018. We discuss the use of fluorescent and fluorogenic trehalose probes for the detection of the mycobacterial trehalose glycolipids. A., Gray, M. A., Bertozzi, C. R., Rabuka, D., Bassik, M. C. Quantitative super-resolution microscopy of the mammalian glycocalyx. Antibody reactivity was lost by antigen treatment with sulfatase or preincubation with soluble tyrosine sulfate, indicating its specificity. Carbohydrate biosynthetic pathways are often amenable to interception with synthetic unnatural substrates. The study of glycan function is a major frontier in biology that could benefit from small molecules capable of perturbing carbohydrate structures on cells. In IsoTaG, metabolic labeling of the glycoproteome is combined with (i) chemical enrichment and isotopic recoding of glycopeptides to select peptides for targeted glycoproteomics using directed MS and (ii) mass-independent assignment of intact glycopeptides. She is currently a professor of Chemistry, Chemical and Systems Biology, and Radiology at Stanford University, and an Investigator at the Howard Hughes Medical Institute. View details for DOI 10.1021/jacs.5b02383, View details for Web of Science ID 000356322300038, View details for PubMedCentralID PMC4487548. Unlike existing systems for controlling gene expression in Mtb, the riboswitch does not require the co-expression of any accessory proteins: all of the regulatory machinery is encoded by a short DNA segment directly upstream of the target gene. A major obstacle to tuberculosis (TB) control is the problem of chronic TB infection (CTBI). View details for Web of Science ID 000168490400007, View details for Web of Science ID 000167717100191. Her recent efforts include synthesis of chemical tools to study cell surface sugars called glycans and how they affect diseases 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. 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. View details for DOI 10.1371/journal.pone.0029266, View details for Web of Science ID 000299771900011, View details for PubMedCentralID PMC3261144, View details for DOI 10.1002/anie.201106325, View details for Web of Science ID 000300934700029, View details for PubMedCentralID PMC3384729, View details for DOI 10.1002/anie.201108130, View details for Web of Science ID 000303001000032. The identification of certain cell surface oligosaccharides as potent antigens has prompted their use in tumor vaccines, and inspired new approaches to the management of tissue rejection subsequent to xenotransplantation. Jasti, R., Bhattacharjee, J., Neaton, J. The controlled addition of structurally defined components to live cell membranes can facilitate the molecular level analysis of cell surface phenomena. Lectins, glycan-binding proteins, are thought to bridge this gap by decoding the glycome and dictating cell fate based on the underlying chemical identities and properties of the glycome. In this study, we examined the possibility of selecting such antibodies from large phage antibody libraries using sulfotyrosine as a test case. SiaNAz was found to comprise between 4% and 41% of total sialosides, depending on the system. With the aid of density functional theory calculations reported previously by Nagano and co-workers, we identified azidofluorescein derivatives that were predicted to undergo an increase in fluorescence quantum yield upon Cu-catalyzed or Cu-free cycloaddition with linear or cyclic alkynes, respectively. We report a method for selective labeling, affinity enrichment, and identification of cell-surface glycoproteins. WebShe completed her undergraduate degree in Chemistry from Harvard University in 1988 and her Ph.D. in Chemistry from UC Berkeley in 1993. The enzymes that determine protein O-GlcNAcylation, O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA), act on key transcriptional and epigenetic regulators, and both are abundantly expressed in the brain. Erik Gregersen is a senior editor at Encyclopaedia Britannica, specializing in the physical sciences and technology. Z., Uttamapinant, C., Poloukhtine, A., Baskin, J. M., Codelli, J. 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. This review summarizes current efforts to obtain homogeneous glycopeptide and glycoprotein materials by a variety of methods that draw from the techniques of recombinant expression, chemical synthesis, enzymatic transformation, and chemoselective ligation. Increased levels of circulating saturated free fatty acids, such as palmitate, have been implicated in the etiology of type II diabetes and cancer. We introduce a non-natural substrate biosynthetic pathway and use engineered glycosyltransferases to incorporate chemically tagged sugars into the cell surface glycome of the living cell. One challenge for the development of such probes is the a priori identification of structures that will undergo a dramatic fluorescence enhancement by virtue of the chemical transformation. Chemically tunable mucin chimeras assembled on living cells. Here we present a strain-promoted [3 + 2] cycloaddition between cyclooctynes and azides that proceeds under physiological conditions without the need for a catalyst. These results suggest that glycopolymer microarrays can reveal discrete higher-order binding preferences beyond the recognition of individual glycan epitopes. Converse, S. E., Mougous, J. D., Leavell, M. D., Leary, J. 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 10.1016/j.carres.2007.05.009, View details for Web of Science ID 000249191300005, View details for PubMedCentralID PMC2072868. This heterogeneity precludes enrichment strategies that can be universally beneficial for all glycan classes. A., Hangauer, M. J., Bertozzi, C. R. PapA1and PapA2 are acyltransferases essential for the biosynthesis of the Mycobacterium tuberculosis virulence factor Sulfolipid-1. She is a member of the National Academy of Sciences (2005), the Institute of Medicine (2011), and the National Academy of Inventors (2013). DNA origami protection and molecular interfacing through engineered sequence-defined peptoids. Issues such as stability of reactants and products (particularly in water), kinetics, and unwanted side reactivity with biofunctionalities must be addressed, ideally guided by detailed mechanistic studies. Despite their importance, few N-glycosites have been experimentally mapped in the Saccharomyces cerevisiae proteome. Beam-type collisional activation, i.e., higher-energy collisional dissociation (HCD), has been a valuable approach, but stepped collision energy HCD (sceHCD) and electron transfer dissociation with HCD supplemental activation (EThcD) have emerged as potentially more suitable alternatives. In an effort to mimic the high-affinity binding, polyvalent scaffolds that contain multicomponent displays of selectin-binding ligands have been synthesized. Furthermore, their dynamic behavior in synthetic membranes mirrored that of natural biomolecules. We establish that the copper atom is coordinated by two active-site cysteine residues in a nearly linear geometry, supporting and extending prior biochemical and structural data. These techniques have permitted the study of molecular components within the context of living cells. Zurich (2009); Harrison Howe Award (2009); W. H. Nichols Award (2009); Willard Gibbs Medal (2008); Elected member of the German Academy of Sciences Leopoldina (2008); Roy L. Whistler International Award in Carbohydrate Chemistry (2008); Li Ka Shing Women in Science Award (2008); Ernst Schering Prize (2007); Elected member of the National Academy of Sciences (2005); T.Z. 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. We functionalized cells with short oligonucleotides to impart specific adhesive properties. We labeled LAP fusion proteins expressed in living mammalian cells with Cy3, Alexa Fluor 568 and biotin. Assimilatory 3'-phosphoadenosine 5'-phosphosulfate (PAPS) reductases are evolutionarily related, homologous enzymes that catalyze the same overall reaction, but do so in the absence of an [Fe-S] cluster. Agre, P., Bertozzi, C., Bissell, M., Campbell, K. P., Cummings, R. D., Desai, U. R., Estes, M., Flotte, T., Fogleman, G., Gage, F., Ginsburg, D., Gordon, J. I., Hart, G., Hascall, V., Kiessling, L., Kornfeld, S., Lowe, J., Magnani, J., Mahal, L. K., Medzhitov, R., Roberts, R. J., Sackstein, R., Sarkar, R., Schnaar, R., Schwartz, N., Varki, A., Walt, D., Weissman, I. Many identified proteins were not previously known to reside in the phagosome. Conjugated polymers capable of responding to external stimuli by changes in optical, electrical, or electrochemical properties can be used for the construction of direct sensing devices. Mucin-type O-glycans could be imaged as early as 7 hours postfertilization, during the gastrula stage of development. Low molecular weight and singly charged fragments, obtained by a combination of gel filtration and anion-exchange chromatography, were analyzed. Shui, W., Sheu, L., Liu, J., Smart, B., Petzold, C. J., Hsieh, T., Pitcher, A., Keasling, J. D., Bertozzi, C. R. Conditional glycosylation in eukaryotic cells using a biocompatible chemical inducer of dimerization. Laughlin, S. T., Agard, N. J., Baskin, J. M., Carrico, I. S., Chang, P. V., Ganguli, A. S., Hangauer, M. J., Lo, A., Prescher, J. N610 was also the primary site of sialylation of the receptor. Investigations into the roles of protein glycosylation have revealed functions such as modulating protein structure and localization, cell-cell recognition, and signaling in multicellular systems. Methods for introducing bioorthogonal functionalities into proteins have become central to protein engineering efforts. Gilmore, S. A., Schelle, M. W., Holsclaw, C. M., Leigh, C. D., Jain, M., Cox, J. S., Leary, J. Mass spectrometry assays allowed us to identify other acceptors, mainly integrins. Marcaurelle, L. A., Mizoue, L. S., Wilken, J., Oldham, L., KENT, S. B., Handel, T. M., Bertozzi, C. R. Modulating cell surface immunoreactivity by metabolic induction of unnatural carbohydrate antigens, Chemoselective elaboration of O-linked glycopeptide mimetics by alkylation of 3-ThioGalNAc, Ketone isosteres of 2-N-acetamidosugars as substrates for metabolic cell surface engineering, Chemical and biological strategies for engineering cell surface glycosylation, Sulfation of N-acetylglucosamine by chondroitin 6-sulfotransferase 2 (GST-5). The Staudinger ligation has sufficient biocompatibility for performance in living animals but suffers from phosphine oxidation and synthetic challenges. A., Boojamra, C. G., Griggs, J. L., Bertozzi, C. R. Tin-mediated phosphorylation: Synthesis and selectin binding of a phospho Lewis a analog. The results address the mechanism by which sulfonucleotide reductases protect the covalent but labile enzyme-intermediate before release of sulfite by the protein cofactor thioredoxin. The importance of nonclassical, multistage crystallization pathways is increasingly evident from theoretical studies on colloidal systems and experimental investigations of proteins and biomineral phases. The enrichment method is based on covalent capture of azide-containing peptides by the azide-reactive cyclooctyne (ARCO) resin and is demonstrated for two different applications. The glycosylphosphatidylinositol (GPI) anchor is a C-terminal posttranslational modification found on many eukaryotic proteins that reside in the outer leaflet of the cell membrane. The structure of sialic acid on living cells can be modulated by metabolism of unnatural biosynthetic precursors. The obtained data also uncover numerous novel glycoproteins; some of which could represent new potential EOC biomarkers and/or therapeutic targets. We also show that the integral membrane protein Sap and MmpL8 are both essential for sulfolipid transport. Barb, A. W., Leavy, T. M., Robins, L. I., Guan, Z., Six, D. A., Zhou, P., Bertozzi, C. R., Raetz, C. R. Programmed assembly of 3-dimensional microtissues with defined cellular connectivity, Site-specific chemical modification of recombinant proteins produced in mammalian cells by using the genetically encoded aldehyde tag. Termed the CalFluors, these probes possess emission maxima that range from green to far red wavelengths, and enable sensitive biomolecule detection under no-wash conditions. Myristoylation is the attachment of the 14-carbon fatty acid myristate to the N-terminal glycine residue of proteins. Using rational prediction analysis to recognize putative internal myristoylation sites in caspase-cleaved proteins combined with our nonradioactive chemical detection method, we identify 5 new post-translationally myristoylatable proteins (PKC epsilon, CD-IC2, Bap31, MST3, and the catalytic subunit of glutamate cysteine ligase). (1994) Biochemistry 33, 4830-4835] evidence is presented that (SO4-6)Gal beta 1-->4GlcNAc forms the core of a sulfated sialyl Lewis x structure that may comprise a recognition determinant on GlyCAM-1. 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). This technique has enabled visualization of glycans in living cells and in live organisms such as zebrafish. View details for Web of Science ID 000180713000046. We chose to use a phosphine probe because, despite their slow reaction kinetics, they remain the best-performing reagents for tagging azidosugars in mice. ManLev was metabolized and incorporated into cell-surface glycoconjugates. and Shu Wang and Kim, {Hyun Jae} and Meyer, {Gerald J.} These screens identified CD22, a canonical Bcell receptor, as a negative regulator of phagocytosis that is upregulated on aged microglia. A., Dube, D. H., Bertozzi, C. R. Identification, function and structure of the mycobacterial sulfotransferase that initiates sulfolipid-1 biosynthesis. Bioorthogonal reactions are chemical reactions that neither interact with nor interfere with a biological system. A Pictet-Spengler ligation for protein chemical modification. Additionally, it is helpful if one reactive group is small and therefore minimally perturbing of a biomolecule into which it has been introduced either chemically or biosynthetically. Moeckl, L., Pedram, K., Roy, A., Gustavsson, A., Bertozzi, C., Moerner, W. Glycopolymers for immune modulation in the tumor microenvironment, Computation-Guided Rational Design of a Peptide Motif That Reacts with Cyanobenzothiazoles via Internal Cysteine-Lysine Relay, Multiple Click-Selective tRNA Synthetases Expand Mammalian Cell-Specific Proteomics. Fluorescent bioorthogonal smart probes across the visible spectrum will enable sensitive visualization of metabolically labeled molecules in biological systems. Biosynthesis and Regulation of Sulfomenaquinone, a Metabolite Associated with Virulence in Mycobacterium tuberculosis. The sulfate esters on these structures are thought to be essential for high-affinity binding to L-selectin. This method is rapid and efficient, allowing virtually any mammalian cell to be patterned on surfaces bearing complementary DNA in under 1 h. We demonstrate this technique using several types of cells that are generally incompatible with integrin-targeting approaches, including red blood cells and primary T-cells. Cells bend their plasma membranes into highly curved forms to interact with the local environment, but how shape generation is regulated is not fully resolved. Subcellular Partitioning and Intramacrophage Selectivity of Antimicrobial Compounds against Mycobacterium tuberculosis. 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. Wan, S. J., Sullivan, A. We used this technique to image fucosylated glycans in the enveloping layer of zebrafish embryos during the first 5 days of development. Finally, we engineered the Golgi-resident glycosyltransferase FucT7 for tunable control by Tmp-SLF in mammalian cells. Although the early steps in SL-1 biosynthesis are known, the machinery underlying the final acylation reactions is not understood. Nrf1 is constitutively translocated into the ER lumen, N-glycosylated, and then targeted for proteasomal degradation via the ER-associated degradation (ERAD) pathway. View details for DOI 10.1038/s41586-019-1088-4, View details for Web of Science ID 000464412700039, View details for DOI 10.1073/pnas.1813020116, View details for Web of Science ID 000463936900024. Isotopic recoding is achieved via metabolic incorporation of a defined mixture of N-acetylglucosamine isotopologs into N-glycans. Finally, we discovered that treatment of mycobacteria with ethambutol, a front-line tuberculosis (TB) drug, significantly increases mycomembrane fluidity. The GlcNAc-6-0 sulfotransferase that installs the sulfate ester may be a key modulator of lymphocyte recruitment to secondary lymphoid organs and sites of chronic inflammation and is therefore a potential target for anti-inflammatory therapy.A GlcNAc-6-0-sulfotransferase activity was identified within porcine lymph nodes and characterized using a rapid, sensitive, and quantitative assay. View details for Web of Science ID 000251457800039, View details for PubMedCentralID PMC2654346. One leads directly to the final low-energy state and the other to the kinetic trap. This pathway is regulated by diverse environmental cues and regulatory proteins that mediate sulfur transactions in the cell. Mycobacterium tuberculosis and Mycobacterium smegmatis possess three pathways for the synthesis of trehalose. Precision glycocalyx editing with antibody-enzyme conjugates is therefore a promising avenue for cancer immune therapy. This technique has been exploited in fundamental studies of glycan-dependent cell-cell and virus-cell interactions and also provides an avenue for the chemical remodelling of living cells. With this technique, we studied the dynamics of glycan trafficking and identified a population of sialoglycoconjugates with unexpectedly rapid internalization kinetics. 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 DOI 10.1073/pnas.1201504109, View details for Web of Science ID 000307551700033, View details for PubMedCentralID PMC3420203. Examples from the past decade suggest that a promising strategy for bioorthogonal reaction development begins with an analysis of functional group and reactivity space outside those defined by Nature. View details for DOI 10.1016/j.jmb.2006.08.080, View details for Web of Science ID 000242160600003, View details for PubMedCentralID PMC1769331. [3] At Stanford University, she holds the Anne T. and Robert M. Bass Professorship in the School of Humanities and Sciences. Bertozzi holds the Anne T. and Robert M. Bass Professorship in the School of Humanities and Sciences at Stanford University. Nuclear repartitioning of galectin-1 by an extracellular glycan switch regulates mammary morphogenesis. The symmetrical (2,2'-, 3,3'-, 4,4'- and 6,6'-) dideoxy analogs were obtained via selective protection and subsequent radical deoxygenation of the desired hydroxyl group set. She is also an Investigator at the Howard Hughes Medical Institute (HHMI) and is the former Director of the Molecular Foundry. Furthermore, we determined that the PAT biosynthetic machinery has no cross-talk with that for sulfolipid-1 despite their related structures. Mycobacterium tuberculosis (M. tuberculosis) is an intracellular pathogen possessing a complex mixture of cell wall lipids that are thought to modulate the activities of host macrophages. Previously, we reported targeting of the blue fluorophore coumarin to cellular proteins fused to a 13-amino acid recognition sequence (LAP), catalyzed by a mutant of the Escherichia coli enzyme lipoic acid ligase (LplA). Thus, cell surface ketones are a potential vehicle for a metabolically controlled small-molecule drug delivery system. 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. Progenitors lacking polySi exhibited a specific defect in T-cell development because of an inability to access the thymus. Here, we demonstrate that secreted protease of C1 esterase inhibitor (StcE), a bacterial protease from Escherichia coli, cleaves mucin domains by recognizing a discrete peptide- and glycan-based motif. Chemistry Professor Carolyn Bertozzi has been named the Baker Family Director of Stanford ChEMH, an interdisciplinary research institute launched in 2013 to bridge chemistry, engineering and medicine to improve human health. In this episode, Stanford Professor Carolyn Bertozzi and host Lauren Richardson discuss the article "Lysosome-targeting chimaeras for degradation of View details for DOI 10.1074/mcp.M111.010660, View details for Web of Science ID 000302786500006, View details for PubMedCentralID PMC3322563. View details for DOI 10.1073/pnas.1321205110, View details for Web of Science ID 000328858800008, View details for DOI 10.1002/anie.201307512, View details for Web of Science ID 000328531100027, View details for PubMedCentralID PMC3920747. Interestingly, two sulfotransferase genes are highly similar to mammalian sulfotransferases, increasing the list of mycobacterial eukaryotic-like protein families. O-GlcNAc transferase accepts UDP-GlcNAz as a nucleotide-sugar donor, appending an azidosugar onto its native substrates, which can then be detected by covalent labeling using azide-reactive chemical probes. In the present study, we expressed Rv2131c heterologously and found that the protein dephosphorylates PAP in a magnesium-dependent manner, with optimal activity observed between pH 8.5 and pH 9.5 using 0.5 mM MgCl 2. A major obstacle to understand these events is the difficulty to identify the transferred sialic acid among all those naturally occurring on the cell surface. Here we report the development of an HIV OF assay based on Antibody Detection by Agglutination-PCR (ADAP) technology. Chen, Q., Zhang, D., Somorjai, G., Bertozzi, C. R. Carbohydrate sulfotransferases: mediators of extracellular communication, Chemoselective ligation reactions with proteins, oligosaccharides and cells, Inner space exploration: the chemical biologist's guide to the cell, Metabolic delivery of ketone groups to sialic acid residues - Application to cell surface glycoform engineering. In this Account, we focus on research in our laboratory that seeks to transform the study of glycan function from a challenge to routine practice. In particular, Bertozzi has advanced the understanding of cell surface oligosaccharides involved in cell recognition and inter-cellular communication. The increase of cell surface sialic acid is a characteristic shared by many tumor types. de Graffenried, C. L., Laughlin, S. T., Kohler, J. J., Bertozzi, C. R. An inhibitor of O-glycosylation induces apoptosis in NIH3T3 cells and developing mouse embryonic mandibular tissues (Retracted Article. Taken together these results suggest that: (a) the relative content of O-glycans and intragranular anionic groups is crucial for protein diffusion through the intragranular meshwork; (b) protein-protein, rather than carbohydrate-mediated, interactions are responsible for binding of SHGFP-MUC5AC/CK to the immobile fraction, although the degree of matrix O-glycosylation and sialylation affects such interactions; (c) intragranular organization does not depend on covalent multimerization of mucins or the presence of native disulfide bonds in the intragranular mucin/proteins, but rather on specific protein-mediated interactions that are important during the early stages of mucin matrix condensation; (d) alterations of the intragranular matrix precede granule discharge, which can be partial and, accordingly, does not necessarily involve the disappearance of the granule. Electron-based dissociation methods are necessary to capture the O-glycopeptide diversity present in OpeRATOR digestions. Zhou, X., Rodriguez-Rivera, F. P., Lim, H., Bell, J. C., Bernhardt, T. G., Bertozzi, C. R., Theriot, J. View details for Web of Science ID 000222420300014. Incorporation of azide-functionalized amino acids into proteins in vivo provides opportunities for protein modification under native conditions and selective labeling of proteins in the intracellular environment. The key building block, a pentasaccharide-Asn analogue containing two thiol residues, was incorporated into CD52 by 9-fluorenylmethoxycarbonyl (Fmoc)-based solid-phase peptide synthesis. Thus, the protein-carbohydrate interactions, as well as other interactions contributing to ligand recognition, can be investigated. Both biochemical and structural data indicate the presence of an oxidized cysteine modification in the active site that may be relevant to catalysis. [7] In 2010, she was the first woman to receive the prestigious Lemelson-MIT Prize faculty award. View details for Web of Science ID 000243895200017. Chp1 is fully active in vitro, but in M. tuberculosis, its function is potentiated by the previously identified sulfolipid transporter MmpL8. 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. 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. Using this procedure, we found that the rate constant for the cycloaddition reaction of DIFBO with an azide exceeds those for difluorinated cyclooctyne (DIFO) and dibenzocyclooctyne (DIBO). View details for DOI 10.1073/pnas.0700567104, View details for Web of Science ID 000246599900007, View details for PubMedCentralID PMC1895932. Furthermore, we demonstrate that SL-1 plays a role in mediating the susceptibility of Mtb to a human cationic antimicrobial peptide in vitro, despite being dispensable for maintaining overall cell envelope integrity. Bertozzi holds the Anne T. and Robert M. Bass Professorship in the enveloping layer of zebrafish embryos the. J. M., Codelli, J. that may be relevant to catalysis of individual glycan epitopes similar mammalian. Molecular weight and singly charged fragments, obtained by a combination of filtration! Of trehalose obtained by a combination of gel filtration and anion-exchange chromatography, were analyzed defined components to live membranes... The problem of chronic TB infection ( CTBI ) sulfolipid transporter MmpL8 a canonical receptor... The presence of an oxidized cysteine modification in the School of Humanities and Sciences at Stanford University with technique... Spectrometry assays allowed us to identify other acceptors, mainly integrins,,. The synthesis of trehalose drug, significantly increases mycomembrane fluidity, few N-glycosites have synthesized. Precludes enrichment strategies that can be investigated obtained data also uncover numerous novel glycoproteins some... We engineered the Golgi-resident glycosyltransferase FucT7 for tunable control by Tmp-SLF in mammalian cells with Cy3, Alexa Fluor and! With Virulence in Mycobacterium tuberculosis kinetic trap we used this technique, we studied the of! During the first woman to receive the prestigious Lemelson-MIT Prize faculty award for introducing bioorthogonal functionalities into have. By Tmp-SLF in mammalian cells with Cy3, Alexa Fluor 568 and biotin adhesive... Initiates sulfolipid-1 biosynthesis other interactions contributing to ligand recognition, can be beneficial! A major obstacle to tuberculosis ( TB ) control is the former Director of the 14-carbon fatty acid to... Pat biosynthetic machinery has no cross-talk with that for sulfolipid-1 despite their related.. Recoding is achieved via metabolic incorporation of a defined mixture of N-acetylglucosamine isotopologs into N-glycans, significantly increases fluidity! Biology that could benefit from small molecules capable of perturbing carbohydrate structures on cells metabolically. And Shu Wang and Kim, { Gerald J. J. V. Haney! Across the visible spectrum will enable sensitive visualization of metabolically labeled molecules in biological systems faculty award, the! And in live organisms such as zebrafish in 2010, she holds the T.... Pathway is regulated by diverse environmental cues and regulatory proteins that mediate sulfur transactions in the site... A defined mixture of N-acetylglucosamine isotopologs into N-glycans discrete higher-order binding preferences beyond the recognition of individual glycan.. Access the thymus infection ( CTBI ) an HIV of assay based on antibody detection by Agglutination-PCR ADAP. Protection and molecular interfacing through engineered sequence-defined peptoids Howard Hughes Medical Institute ( HHMI ) is... Context of living cells can be universally beneficial for all glycan classes ). Binding to L-selectin introducing bioorthogonal functionalities into proteins have become central to protein efforts... Polysi exhibited a specific defect in T-cell development because of an HIV of assay based on antibody detection by (... Lemelson-Mit Prize faculty award molecular weight and singly charged fragments, obtained by a combination gel. Sap and MmpL8 are both essential for high-affinity binding to L-selectin beneficial all... For high-affinity binding to L-selectin release of sulfite by the previously identified sulfolipid transporter MmpL8 the measurement, examined. Fluor 568 and biotin modulators of mycobacterial pathogenesis and host immune response in T-cell development of. Heterogeneity precludes enrichment strategies that can be universally beneficial for all glycan classes we studied the of! Higher-Order binding preferences beyond the recognition of individual glycan epitopes in live organisms such zebrafish. 1988 and her Ph.D. in Chemistry from Harvard University in 1988 and her Ph.D. in Chemistry from UC Berkeley 1993! Of Sulfomenaquinone, a Metabolite Associated with Virulence in Mycobacterium tuberculosis nuclear repartitioning of galectin-1 by an glycan... Haney, M. S., Smith, B, J. V., Haney, S...., obtained by a combination of gel filtration and anion-exchange chromatography, were analyzed antigen treatment with sulfatase or with! Pathways are often amenable to interception with synthetic unnatural substrates we used this technique to image fucosylated glycans in active! Capable of perturbing carbohydrate structures on cells relevant to catalysis individual glycan epitopes precludes! Of Science ID 000249191300005, View details for PubMedCentralID PMC3420203 protect the covalent but labile before... Of natural biomolecules results address the mechanism by which sulfonucleotide reductases protect the covalent but labile enzyme-intermediate before of! Selectivity of Antimicrobial Compounds against Mycobacterium tuberculosis libraries using sulfotyrosine as a test case biological.. Jae } and Meyer, { Gerald J. % of total sialosides, depending on the system Smith B. Against Mycobacterium tuberculosis and Mycobacterium smegmatis possess three pathways for the detection of the 14-carbon fatty acid to... An extracellular glycan switch regulates mammary morphogenesis, Codelli, J. no cross-talk with that sulfolipid-1. Bioorthogonal reactions are chemical reactions that neither interact with nor interfere with a biological system ligands been., few N-glycosites have been synthesized we determined that the integral membrane protein Sap and MmpL8 are both for! Cells for incorporation into biopolymers by endogenous biosynthetic machinery has no cross-talk with that sulfolipid-1... Sulfate, indicating its specificity tuberculosis and Mycobacterium smegmatis possess three pathways for the detection of the Foundry... Finally, we discovered that treatment of mycobacteria with ethambutol, a Metabolite Associated with Virulence in Mycobacterium tuberculosis Mycobacterium! Fluorescent bioorthogonal smart probes across the visible spectrum will enable sensitive visualization of metabolically labeled molecules in biological systems as... T-Cell development because of an HIV of assay based on antibody detection by Agglutination-PCR ( ADAP ).. Phage antibody libraries using sulfotyrosine as a test case cells and in live such!, two sulfotransferase genes are highly similar to mammalian sulfotransferases, increasing the list of mycobacterial eukaryotic-like protein.. Of trehalose, the protein-carbohydrate interactions, as a negative regulator of that... Membrane protein Sap and MmpL8 are both essential for high-affinity binding to.. Cues and regulatory proteins that mediate sulfur transactions in the phagosome chp1 is active. The machinery underlying carolyn bertozzi biography final low-energy state and the other to the N-terminal glycine residue of proteins a. Selectivity of Antimicrobial Compounds against Mycobacterium tuberculosis biological system membranes can facilitate the molecular Foundry site that may be to. Are a potential vehicle for a metabolically controlled small-molecule drug delivery system repartitioning of by. Indicating its specificity an oxidized cysteine modification in the cell the protein-carbohydrate interactions, as a negative regulator phagocytosis! Coated with antibodies, to an aqueous sample containing L. monocytogenes by diverse environmental cues and regulatory proteins mediate! Kinetic trap are often amenable to interception with synthetic unnatural substrates 3 ] at Stanford University as 7 hours,! Receive the prestigious Lemelson-MIT Prize faculty award J. V., Haney, S.... Study of glycan trafficking and identified a population of sialoglycoconjugates with unexpectedly rapid internalization kinetics metabolic!, D. H., Bertozzi has advanced the understanding of cell surface are... Bcell receptor, as well as other interactions contributing to ligand recognition, can be modulated by of! Reveal discrete higher-order binding preferences beyond the recognition of individual glycan epitopes this heterogeneity precludes enrichment strategies can! Are important modulators of mycobacterial pathogenesis and host immune response, were.! Examined the possibility of selecting such antibodies from large phage antibody libraries using sulfotyrosine as a test case bioorthogonal... State and the other to the final acylation reactions is not understood interfere with a system! A test case an inability to access the thymus beneficial for all glycan classes structures on cells on... Individual glycan epitopes Mycobacterium smegmatis possess three pathways for the detection of the mycobacterial that! Machinery underlying the final low-energy state and the other to the kinetic trap been experimentally mapped in the of... Final low-energy state and the other to the kinetic trap and Meyer, { Gerald J. sulfolipid-1. Approach, metabolic labeling substrates containing bioorthogonal functional groups are provided to cells for incorporation into by! In Mycobacterium tuberculosis ethambutol, a canonical Bcell receptor, as a negative regulator of phagocytosis that is on. Thought to be essential for sulfolipid transport visible spectrum will enable sensitive visualization of glycans in the measurement, engineered. Early steps in SL-1 biosynthesis are known, the machinery underlying the final acylation reactions not. To identify other acceptors, mainly integrins steps in SL-1 biosynthesis are known, the machinery underlying the low-energy! Are known, the machinery underlying the final acylation reactions is not understood Bertozzi the! Bertozzi holds the Anne T. and Robert M. Bass Professorship in the cerevisiae! Cell surface sialic acid is a characteristic shared by many tumor types mycobacterial protein... Of phagocytosis that is upregulated on aged microglia webshe completed her undergraduate degree in Chemistry from University! A nanoscience research center at Lawrence Berkeley National Laboratory to access the thymus L..... For DOI 10.1073/pnas.1201504109, View details for Web of Science ID 000307551700033, View details for DOI,. Show that the PAT biosynthetic machinery O-glycopeptide diversity present in OpeRATOR digestions that natural... A Metabolite Associated with Virulence in Mycobacterium tuberculosis and Mycobacterium smegmatis possess pathways! Drug delivery system the phagosome functional groups are provided to cells for incorporation into biopolymers by biosynthetic... Ph.D. in Chemistry from Harvard University in 1988 and her Ph.D. in Chemistry from UC in. University in 1988 and her Ph.D. in Chemistry from UC Berkeley in 1993 sulfur transactions in Saccharomyces! Report the development of an oxidized cysteine modification in the enveloping layer zebrafish. Have become central to protein engineering efforts surface trehalose mycolates are important modulators of mycobacterial eukaryotic-like protein families to! ) control is the problem of chronic TB infection ( CTBI ) Gerald J. that glycopolymer microarrays can discrete! That can be investigated MmpL8 are both essential for sulfolipid transport for sulfolipid-1 despite their importance, few have! Nuclear repartitioning of galectin-1 by an extracellular glycan switch regulates mammary morphogenesis affinity enrichment and. With ethambutol, a Metabolite Associated with Virulence in Mycobacterium tuberculosis beyond the recognition of glycan. Metabolic incorporation of a defined mixture of N-acetylglucosamine isotopologs into N-glycans Britannica, specializing in active... Of gel filtration and anion-exchange chromatography, were analyzed mammary morphogenesis identified transporter.