A5044077084- Glycosylation and Glycoproteins Research
- Carbohydrate Chemistry and Synthesis
- Advanced Proteomics Techniques and Applications
- Monoclonal and Polyclonal Antibodies Research
- Genomics and Phylogenetic Studies
- Immunotherapy and Immune Responses
- Galectins and Cancer Biology
- Immune Cell Function and Interaction
- Ubiquitin and proteasome pathways
- Peptidase Inhibition and Analysis
- vaccines and immunoinformatics approaches
- Cancer Research and Treatments
- T-cell and B-cell Immunology
- RNA and protein synthesis mechanisms
- Enzyme Production and Characterization
- Mass Spectrometry Techniques and Applications
- Click Chemistry and Applications
- Viral Infectious Diseases and Gene Expression in Insects
- Microbial Metabolites in Food Biotechnology
- Reproductive System and Pregnancy
- Bacteriophages and microbial interactions
- Virus-based gene therapy research
- Platelet Disorders and Treatments
- Cancer Immunotherapy and Biomarkers
- Tryptophan and brain disorders
University of Virginia
2013-2025
Yale University
2020-2025
Stanford University
2017-2023
Protéomique, Réponse Inflammatoire et Spectrométrie de Masse
2020-2021
Inserm
2020-2021
Université de Lille
2020-2021
Howard Hughes Medical Institute
2020
Mucin domains are densely O-glycosylated modular protein that found in a wide variety of cell surface and secreted proteins. Mucin-domain glycoproteins known to be key players host human diseases, especially cancer, wherein mucin expression glycosylation patterns altered. biology has been difficult study at the molecular level, part, because methods manipulate structurally characterize lacking. Here, we demonstrate protease C1 esterase inhibitor (StcE), bacterial from Escherichia coli,...
Site-specific characterization of glycosylation requires intact glycopeptide analysis, and recent efforts have focused on how to best interrogate glycopeptides using tandem mass spectrometry (MS/MS). Beam-type collisional activation, i.e., higher-energy dissociation (HCD), has been a valuable approach, but stepped collision energy HCD (sceHCD) electron transfer with supplemental activation (EThcD) emerged as potentially more suitable alternatives. Both sceHCD EThcD used success in...
Abstract Mucin domains are densely O-glycosylated modular protein found in various extracellular and transmembrane proteins. Mucin-domain glycoproteins play important roles many human diseases, such as cancer cystic fibrosis, but the scope of mucinome remains poorly defined. Recently, we characterized a bacterial O-glycoprotease, StcE, demonstrated that an inactive point mutant retains binding selectivity for mucin-domain glycoproteins. In this work, leverage StcE to selectively enrich...
Targeted protein degradation is an emerging strategy for the elimination of classically undruggable proteins. Here, to expand landscape targetable substrates, we designed degraders that achieve substrate selectivity via recognition a discrete peptide and glycan motif cell-type antigen-driven cell-surface binding. We applied this approach mucins, O-glycosylated proteins drive cancer progression through biophysical immunological mechanisms. Engineering bacterial mucin-selective protease...
Immunity against phosphopeptide antigens lacking in leukemia patients can be restored with stem cell transplantation.
Leukemias are highly immunogenic, but they have a low mutational load, providing few mutated peptide targets. Thus, the identification of alternative neoantigens is pressing need. Here, we identify 36 MHC class I-associated antigens with O-linked β-N-acetylglucosamine (O-GlcNAc) modifications as candidate neoantigens, using three experimental approaches. Thirteen these peptides were also detected disaccharide units on same residues and two contain either mono- and/or di-methylated arginine...
Different approaches are used in the production of recombinant adeno-associated virus (rAAV). The two leading transiently transfected human HEK293 cells and live baculovirus infection Spodoptera frugiperda (Sf9) insect cells. Unexplained differences vector performance have been seen clinically preclinically. Thus, we performed a controlled comparative analysis varying only host cell species but maintaining all other parameters. We characterized with multiple analytical approaches: proteomic...
Significance Cancer cells are frequently coated with chains of sugar molecules (glycans) that bind to inhibitory immune receptors, leading suppression anticancer immunity. While blocking these interactions may be therapeutically beneficial, the specific glycoprotein ligands engage such receptors often complex and difficult characterize. To solve this problem, we used unbiased genome-wide screening identify genes required for cell-surface presentation members Siglec receptor family. This...
Studying posttranslational modifications classically relies on experimental strategies that oversimplify the complex biosynthetic machineries of living cells. Protein glycosylation contributes to essential biological processes, but correlating glycan structure, underlying protein, and disease-relevant regulation is currently elusive. Here, we engineer cells tag glycans with editable chemical functionalities while providing information biosynthesis, physiological context, fine structure. We...
Densely O-glycosylated mucin domains are found in a broad range of cell surface and secreted proteins, where they play key physiological roles. In addition, alterations expression glycosylation common variety human diseases, such as cancer, cystic fibrosis, inflammatory bowel diseases. These correlations have been challenging to uncover establish because tools that specifically probe lacking. Here, we present panel bacterial proteases cleave via distinct peptide- glycan-based motifs,...
Significance Most human secreted and cell surface proteins are modified by Ser/Thr(O)-linked glycosylation with N -acetylgalactosamine (O-GalNAc). While of fundamental importance in health disease, O-GalNAcglycosylation is technically challenging to study because a lack specific tools for biological assays. Here, we design an O-GalNAc–specific reporter molecule termed uridine diphosphate (UDP)– -( S )-azidopropionylgalactosamine (GalNAzMe) selectively label O-GalNAc glycoproteins living...
Abstract Mucin-domain glycoproteins are densely O-glycosylated and play critical roles in a host of biological functions. In particular, the T cell immunoglobulin mucin-domain containing family proteins (TIM-1, -3, -4) decorate immune cells act as key regulators cellular immunity. However, their dense O-glycosylation remains enigmatic, primarily due to challenges associated with studying mucin domains. Here, we demonstrate that mucinase SmE has unique ability cleave at residues bearing very...
O-Linked α-N-acetylgalactosamine (O-GalNAc) glycans constitute a major part of the human glycome. They are difficult to study because complex interplay 20 distinct glycosyltransferase isoenzymes that initiate this form glycosylation, polypeptide N-acetylgalactosaminyltransferases (GalNAc-Ts). Despite proven disease relevance, correlating activity individual GalNAc-Ts with biological function remains challenging due lack tools probe their substrate specificity in environment. Here, we develop...
Epithelial surfaces throughout the body are coated by mucins, a class of proteins carrying domains characterized high density O-glycosylated serine and threonine residues. The resulting mucosal layers form crucial host-microbe interfaces that prevent translocation microbes while also selecting for distinct bacteria via presented glycan repertoire. intricate interplay between mucus production breakdown thus determines composition microbiota maintained within these environments, which can have...
Abstract Altered glycoprotein expression is an undisputed corollary of cancer development. Understanding these alterations paramount but hampered by limitations underlying cellular model systems. For instance, the intricate interactions between tumour and host cannot be adequately recapitulated in monoculture tumour-derived cell lines. More complex co-culture models usually rely on sorting procedures for proteome analyses rarely capture details protein glycosylation. Here, we report a...
The emergence of a polybasic cleavage motif for the protease furin in SARS-CoV-2 spike has been established as major factor human viral transmission. region N-terminal to that is extensively mutated variants concern (VOCs). Besides furin, spikes from these appear rely on other proteases maturation, including TMPRSS2. Glycans near site have raised questions about proteolytic processing and consequences variant-borne mutations. Here, we identify sialic acid-containing O-linked glycans Thr678...
Abstract Bacterial biofilms are formed on environmental surfaces and host tissues, facilitate colonization antibiotic resistance by human pathogens. Bacteria often express multiple adhesive proteins (adhesins), but it is unclear whether adhesins have specialized or redundant roles. Here, we show how the model biofilm-forming organism Vibrio cholerae uses two with overlapping distinct functions to achieve robust adhesion diverse surfaces. Both biofilm-specific Bap1 RbmC function as a...
Abstract Glycosylation is an incredibly common and diverse post‐translational modification that contributes widely to cellular health disease. Mass spectrometry the premier technique study glycoproteins; however, glycoproteomics has lagged behind traditional proteomics due challenges associated with studying glycosylation. For instance, glycans dissociate by collision‐based fragmentation, thus necessitating electron‐based fragmentation for site‐localization. The vast glycan heterogeneity...
The MHC class II (MHCII) processing pathway presents peptides derived from exogenous or membrane-bound proteins to CD4+ T cells. Several studies have shown that glycopeptides are necessary modulate cell recognition, though glycopeptide structures in these cases generally unknown. Here, we present a total of 93 three melanoma lines and one matched EBV-transformed line with most found only the lines. glycosylation detected was diverse comprised 17 different glycoforms. We then used molecular...