O-glycosylation Analysis Service

Protein O-glycosylation is a common glycosylation modification that plays a key role in tissue formation and is involved in a variety of life activities such as cell recognition, immune response, and cell differentiation. Creative Proteomics offers high-quality O-glycosylation analysis services, including identification, quantification and characterization, providing valuable insights into your protein research.

What is O-glycosylation

Protein O-glycosylation is an intricate post-translational modification where sugar molecules attach to the oxygen atoms of specific serine or threonine residues in a protein. In contrast to N-glycosylation, which involves the en bloc transfer of preassembled oligosaccharides, O-glycosylation is characterized by the stepwise addition of sugars to the protein backbone. This modification predominantly takes place in the Golgi apparatus and plays a role in regulating various cellular processes.

Fig.1 Schematic of O-glycosylation (Gonzalez-Morelo, K. J.; et al. 2020)

The initiation of O-glycosylation involves the addition of a single N-acetylgalactosamine (GalNAc) to the hydroxyl group of a serine or threonine residue, forming the Tn antigen. Subsequent elongation of the O-glycan chain includes the addition of different sugar residues, resulting in diverse glycan structures. Unlike N-glycosylation, O-glycosylation is structurally more varied and diverse, which contributes to the diversity of functions of O-glycosylated proteins. It influences protein folding, stability, and interactions with other molecules in cellular life processes.

Applications of O-glycosylation Analysis

• Cell Signaling Study
  Protein O-glycosylation plays pivotal roles in diverse physiological processes, contributing to cell adhesion, immune response, and the organization of the extracellular matrix.
• Disease Research
  Dysregulation of O-glycosylation is linked to various diseases, and its changes in structure can function as biomarkers, aiding in diagnosis.
• Drug Development
  In the realm of drug development, the engineering of O-glycosylation is a critical consideration, especially in the production of therapeutic proteins like monoclonal antibodies. Ensuring proper O-glycosylation is essential to guarantee the efficacy and safety of these therapeutic interventions.
• Therapeutic Development
  A comprehensive understanding of protein O-glycosylation principles is crucial for unraveling diseases' molecular mechanisms and developing targeted therapeutic interventions.

Our Service

Creative Proteomics has advanced protein detection technologies and can provide customized services to researchers and scientists to help them reveal the complexity of PTM using powerful mass spectrometry technologies. Our services are thoughtful and detailed and the process of O-glycosylation analysis service is as follows:

Fig.2 O-glycosylation analysis workflow

• Sample Preparation
  Proper extraction and preparation of proteins from biological samples to ensure preservation of O-glycosylated proteins.
• glycosylation Enrichment
  Enrichment techniques are employed to isolate O-glycosylated proteins or peptides. This step is crucial as O-glycosylation modifications are often present in low abundance compared to unmodified proteins.
• Detection
  WB, protein microarrays and LC-MS/MS are powerful tools for the identification and quantification of O-glycosylated peptides, and they provide detection information that facilitates a comprehensive analysis of O-glycosylation.
• Bioinformatics Analysis
  Advanced bioinformatics tools are used to analyze data, identify O-glycosylated peptides, and map these modifications to protein sequences.
• Result Delivery
  After professional data analysis, we deliver results quickly and consistently.

Creative Proteomics brings a wealth of experience to the field of protein research. Our comprehensive services and products cover the entire spectrum of utilizing suitable and efficient gateways. Utilizing our proprietary platform, we have successfully implemented many proteomics research projects. If you are interested, please contact us for more information and a quote.

Reference

• Gonzalez-Morelo, K. J.; et al. Molecular Insights Into O-Linked Glycan Utilization by Gut Microbes. Frontiers in Microbiology. 2020, 11: 591568