Glycosylation structure characterization and allergenicity analysis of sarcoplasmic calcium-binding protein from Litopenaeus vannamei

• The detailed structures of N-glycans conjected to SCP were revealed. • Glycosylation increased the allergenicity of SCP. • Seven linear epitopes and two glycosylated conformational epitopes of SCP were identified. Glycosylation is a post-transcriptional modification that can influence protein structure and function, yet the glycosylation of the major shellfish allergen, sarcoplasmic calcium-binding protein (SCP), remains poorly understood. In this study, we characterized the glycosylation profile of SCP and explored the role of N-glycans in its allergenicity. Tandem mass spectrometry (MS n ) revealed seven distinct N-glycans conjugated to SCP and provided detailed structural identification. ELISA and mast cell degranulation assay using sera from allergic individuals demonstrated that SCP exhibits significant allergenicity, which was markedly reduced following deglycosylation. Additionally, through the integration of various bioinformatics approaches, seven potential epitopes of SCP were predicted, and two glycosylated conformational epitopes were identified. In summary, this study elucidated the detailed glycosylation pattern and allergenic properties of SCP, offering a theoretical foundation for future research on the impact of glycosylation on allergenicity and contributing to the development of hypoallergenic products.