A5088099956- Click Chemistry and Applications
- Glycosylation and Glycoproteins Research
- Chemical Synthesis and Analysis
- Carbohydrate Chemistry and Synthesis
- Peptidase Inhibition and Analysis
- Bacteriophages and microbial interactions
- interferon and immune responses
- Immune Response and Inflammation
- Ubiquitin and proteasome pathways
- RNA and protein synthesis mechanisms
- Gut microbiota and health
- Cancer Research and Treatments
- Probiotics and Fermented Foods
- Antimicrobial Resistance in Staphylococcus
- Monoclonal and Polyclonal Antibodies Research
- Advanced Proteomics Techniques and Applications
- Receptor Mechanisms and Signaling
- Antimicrobial Peptides and Activities
- Clostridium difficile and Clostridium perfringens research
- Antibiotic Resistance in Bacteria
- Adenosine and Purinergic Signaling
- Bacterial Genetics and Biotechnology
- Galectins and Cancer Biology
- Influenza Virus Research Studies
- Endoplasmic Reticulum Stress and Disease
Scripps (United States)
2020-2025
Scripps Institution of Oceanography
2020-2025
Harvard University
2004-2024
University of California, San Francisco
2024
Scripps Research Institute
2021-2024
Rockefeller University
2013-2023
Torrey Pines Institute For Molecular Studies
2023
The Herbert Wertheim UF Scripps Institute for Biomedical Innovation & Technology
2021
The University of Texas Southwestern Medical Center
2012
Stony Brook University
2010
The glycosylation of serine and threonine residues with a single GlcNAc moiety is dynamic posttranslational modification many nuclear cytoplasmic proteins. We describe chemical strategy directed toward identifying O -GlcNAc-modified proteins from living cells or modified in vitro . demonstrate, , that each enzyme the hexosamine salvage pathway, enzymes affect this (UDP-GlcNAc:polypeptidtyltransferase -GlcNAcase), tolerate analogues their natural substrates which N -acyl side chain has been...
Mucin-type O-linked glycoproteins are involved in a variety of biological interactions higher eukaryotes. The biosynthesis these is initiated by family polypeptide N -acetyl-α-galactosaminyltransferases (ppGalNAcTs) that modify proteins the secretory pathway. lack defined consensus sequence for ppGalNAcTs makes prediction mucin-type glycosylation difficult based on primary alone. Herein we present method labeling with unique chemical tag, azide, which permits their selective covalent...
Fatty-acylation of proteins in eukaryotes is associated with many fundamental cellular processes but has been challenging to study due limited tools for rapid and robust detection protein fatty-acylation cells. The development azido-fatty acids enabled the nonradioactive fatty-acylated mammalian cells using Staudinger ligation biotinylated phosphine reagents. However, visualization streptavidin blotting highly variable not ideal proteins. Here we report alkynyl-fatty acid chemical reporters...
The structure of sialic acid on living cells can be modulated by metabolism unnatural biosynthetic precursors. Here we investigate the conversion a panel azide-functionalized mannosamine and glucosamine derivatives into cell-surface sialosides. A key tool in this study is Staudinger ligation, highly selective reaction between modified triarylphosphines azides that produces an amide-linked product. preliminary mechanism reaction, refined conditions for its vivo execution, are reported....
The dynamic modification of nuclear and cytoplasmic proteins by the monosaccharide N -acetyl-glucosamine (GlcNAc) continues to emerge as an important regulator many biological processes. Herein we describe development alkynyl-modified GlcNAc analog (GlcNAlk) a new chemical reporter O-GlcNAc in living cells. This strategy is based on metabolic incorporation reactive functionality into biosynthetic pathway. When combined with Cu(I)-catalyzed [3 + 2] azide-alkyne cycloaddition, this allowed for...
The attachment of lipids onto proteins modulates the activity in many biological settings. analysis protein lipidation, however, is challenging due to relatively few methods for detection lipid-modified proteins. Here we describe synthesis ω-azido-fatty acids as non-radioactive chemical probes rapid visualization fatty-acylated mammalian cells. Following metabolic installation target by cellular enzymes, are selectively biotinylated with a phosphine−biotin reagent via Staudinger ligation and...
The interferon (IFN)-induced transmembrane protein 3 (IFITM3) is a cellular restriction factor that inhibits infection by influenza virus and many other pathogenic viruses. IFITM3 prevents endocytosed particles from accessing the host cytoplasm although little known regarding its regulatory mechanisms. Here we demonstrate localization to antiviral remodeling of endolysosomes differentially regulated S-palmitoylation lysine ubiquitination. Although enhances membrane affinity activity,...
Protein acetylation is a key post-translational modification that regulates diverse biological activities in eukaryotes. Here we report bioorthogonal chemical reporters enable direct in-gel fluorescent visualization and proteome-wide identification of acetylated proteins via Cu(I)-catalyzed azide−alkyne cycloaddition, often termed "click chemistry". We demonstrate two alkynyl-acetyl-CoA analogues, 4-pentynoyl-CoA 5-hexynoyl-CoA, function as efficient substrates lysine acetyltransferase p300...
Significance Cysteine fatty acylation (S-fatty acylation) regulates the stability, trafficking, and activity of proteins in eukaryotes. Current methods to study S-fatty rely on metabolic incorporation acid analogs or selective chemical labeling affinity purification, which limits detection endogenous acylated protein isoforms levels. Here we demonstrate that biochemical exchange acyl groups cysteines with defined mass-tags enables direct visualization The application this mass-tag method...
Protein lipidation and lipid trafficking control many key biological functions in all kingdoms of life. The discovery diverse species their covalent attachment to proteins has revealed a complex regulated network membranes lipidated that are central fundamental aspects physiology human disease. Given the complexity protein targeting mechanisms involved with membrane lipids, precise sensitive methods needed monitor identify these hydrophobic molecules bacteria, yeast, higher...
The intestinal microbiome modulates host susceptibility to enteric pathogens, but the specific protective factors and mechanisms of individual bacterial species are not fully characterized. We show that secreted antigen A (SagA) from Enterococcus faecium is sufficient protect Caenorhabditis elegans against Salmonella pathogenesis by promoting pathogen tolerance. NlpC/p60 peptidoglycan hydrolase activity SagA required generates muramyl-peptide fragments C. in a tol-1-dependent manner. can...
Bioorthogonal chemical reporters are useful tools for visualizing and identifying post-translational modifications on proteins. Here we report the proteomic analysis of mammalian proteins targeted by a series fatty acid ranging from myristic to stearic acid. The large-scale total cell lysates fully solubilized Jurkat T cells identified known fatty-acylated many new candidates, including nuclear in particular histone H3 variants. We demonstrate that histones H3.1, H3.2, H3.3 modified with...
Traditional Chinese Medicines (TCMs) have been historically used to treat bacterial infections. However, the molecules responsible for these anti-infective properties and their potential mechanisms of action remained elusive. Using a high-throughput assay type III protein secretion in Salmonella enterica serovar Typhimurium, we discovered that several TCMs can attenuate this key virulence pathway without affecting growth. Among active TCMs, baicalein, specific flavonoid from Scutellaria...
S-prenylation is an important lipid modification that targets proteins to membranes for cell signaling and vesicle trafficking in eukaryotes. As S-prenylated are often key effectors oncogenesis, congenital disorders, microbial pathogenesis, robust proteomic methods still needed biochemically characterize these lipidated specific types disease states. Here, we report bioorthogonal proteomics of macrophages with improved alkyne-isoprenoid chemical reporter enables large-scale profiling...
Over the past years, fluorescent proteins (e.g., green proteins) have been widely utilized to visualize recombinant protein expression and localization in live cells. Although powerful, tags are limited by their relatively large sizes potential perturbation function. Alternatively, site-specific labeling of with small-molecule organic fluorophores using bioorthogonal chemistry may provide a more precise less perturbing method. This approach involves incorporation unnatural amino acids (UAAs)...
We discovered that Enterococcus faecium (E. faecium), a ubiquitous commensal bacterium, and its secreted peptidoglycan hydrolase (SagA) were sufficient to enhance intestinal barrier function pathogen tolerance, but the precise biochemical mechanism was unknown. Here we show E. has unique composition remodeling activity through SagA, which generates smaller muropeptides more effectively activates nucleotide-binding oligomerization domain-containing protein 2 (NOD2) in mammalian cells. Our...