Decoding Protein Glycosylation by an Integrative Mass Spectrometry-Based De Novo Sequencing Strategy.

ABSTRACT: Glycoproteins, representing more than 50% of human proteins and most biopharmaceuticals, are crucial for regulating various biological processes. The complexity of multiple glycosylation sites often leads to incomplete sequence coverage and ambiguous glycan modification profiles. Here, we developed an integrative mass spectrometry-based approach for decoding unknown glycoproteins, which is featured with the combination of deglycosylation-mediated de novo sequencing with glycosylation site characterization. We utilized the enzymatic deglycosylation of N-/ O-glycans to achieve comprehensive sequence coverage. Additionally, EThcD fragmentation enables the identification of high-quality long peptides, facilitating precise protein assembly. We subsequently applied this meth

SUBMITTER: Gao J 

PROVIDER: S-EPMC11863158 | biostudies-literature | 2025 Feb

REPOSITORIES: biostudies-literature