Premium
Identification and characterization of glycosylation sites on Litopenaeus vannamei hemocyanin
Author(s) -
Zhang Zehui,
Li Ruiwei,
Aweya Jude Juventus,
Wang Fan,
Zhong Mingqi,
Zhang Yueling
Publication year - 2019
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1002/1873-3468.13367
Subject(s) - hemocyanin , litopenaeus , glycosylation , biology , biochemistry , shrimp , concanavalin a , protein subunit , glycoconjugate , microbiology and biotechnology , antibody , immunology , ecology , gene , in vitro
The respiratory glycoprotein hemocyanin has been implicated in immune‐related functions. Using lectin blotting, we show that the binding of shrimp ( Litopenaeus vannamei ) hemocyanin to concanavalin A decreases markedly with O‐glycosidase treatment but not with PNG ase F. Twelve O‐glycosylation sites, three on the large hemocyanin subunit and nine on the small hemocyanin subunit ( HMC s), were identified by LC ‐ MS / MS . Importantly, when the glycosylation sites at Thr‐537, Ser‐539, and Thr‐542 on the C terminus of HMC s were replaced with alanine, the resultant mutant hemocyanin had reduced carbohydrate content, coupled with a fourfold reduction in bacterial agglutination and 0.2‐fold reduction in antibacterial activities toward Vibrio parahaemolyticus and Staphylococcus aureus . These results suggest that the glycosylation sites on shrimp hemocyanin are closely related to its immunological functions.