O-Glycosylation Site Analysis of Escherichia Coli

In today's biological science research, a deep understanding of post-translational modifications of proteins, especially glycosylation, is critically important in revealing protein function, cell signal transduction, pathogen recognition, and disease onset mechanisms. O-glycosylation, as a common post-translational modification, involves the addition of mono- or polysaccharide chains to serine or threonine residues on proteins. In microbes like Escherichia coli, O-glycosylation plays a decisive role in their biological function, pathogenicity, and interactions with the host.

 

In the fields of microbiology and biotechnology, research on bacterial surface glycosylation is key to understanding bacterial physiology and pathogenic mechanisms. Escherichia coli (E. coli), as a widely studied model microbe, its surface O-glycosylation (O-antigen glycosylation) site analysis provides researchers with an important window to understand the interactions between bacteria and hosts, pathogenicity, and mechanisms of immune evasion. MtoZ Biolabs utilizes advanced biotechnology and analysis platforms in this field to provide professional services for O-glycosylation site analysis of E. coli. Through our analysis, researchers can accurately identify the O-glycosylation modification sites of E. coli proteins, gaining a deeper understanding of their biological functions and roles in host interactions.

 

Services at MtoZ Biolabs

1. Glycan Analysis

(1) N-glycans are released with PNGase F and removed from the solution.

(2) Enzymes are used to digest the protein.

(3) O-glycopeptides are released by β-elimination.

(4) O-glycopeptides are enriched with HILIC (Hydrophilic Interaction Liquid Chromatography).

(5) The occupancy rate of O-linked glycosylation sites is determined by LC-ESI-MS/MS.

 

2. Bottom-Up Approach

(1) Digestion

The protein is digested with a protease (usually trypsin) after reduction and alkylation. In addition, we will also use multiple proteases for enzymatic digestion to increase the coverage of O-glycan sites.

 

(2) Enrichment

Amide-80, MAX, or SAX SPE are used for enrichment.

 

(3) Detection

Glycopeptides are directly analyzed by LC-MS/MS.

 

Service Advantages

1. High Precision Site Identification

By using high-resolution mass spectrometry technology combined with specialized enzymatic digestion and enrichment strategies, we can identify O-glycosylation sites with high precision, not missing even low-abundance modifications.

 

2. Wide Coverage

Our technology can not only analyze known O-glycosylation sites but also discover new modification sites, providing a new perspective for the study of E. coli protein function.

 

3. In-Depth Functional Analysis

In addition to the identification of O-glycosylation sites, we also provide quantitative analysis of modification sites, protein-protein interaction analysis, and evaluation of the impact of modifications on protein structure and function, helping clients understand the biological significance of O-glycosylation from multiple dimensions.

 

4. Customized Solutions

Based on the specific research needs of clients, we can provide customized services from sample preparation to data analysis, ensuring accurate, reliable, and biologically meaningful results.

 

MtoZ Biolabs' O-glycosylation site analysis service is based on a profound understanding of biology and the application of advanced technologies, aiming to provide clients with accurate and comprehensive analysis results. We warmly welcome scientists and researchers worldwide to collaborate with us. Feel free to contact us for consultation.