Purification, Characterization, cDNA Cloning, and Bioinformatic Analysis of Zinc-Binding Protein from Magallana hongkongensis

• Published in: Molecules
• Main Authors: Citing Chen, Wan Li, Jialong Gao, Wenhong Cao, Xiaoming Qin, Huina Zheng, Haisheng Lin, Zhongqin Chen
• Format: Article in Journal/Newspaper
• Language: English
• Published: MDPI AG 2024
• Subjects: Magallana hongkongensis; zinc-binding protein; cDNA cloning; carbonic anhydrase; Organic chemistry; QD241-441; Crassostrea gigas
• Online Access: https://doi.org/10.3390/molecules29040900; https://doaj.org/article/0bcea077420049a4b6415e2dd3dc464b

Oysters contain significant amounts of the zinc element, which may also be found in their proteins. In this study, a novel zinc-binding protein was purified from the mantle of the oyster Magallana hongkongensis using two kinds of gel filtration chromatograms. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) showed that its molecular weight was approximately 36 kDa. The protein identified by the Q-Exactive mass spectrometer shared the highest sequence identity with carbonic anhydrase derived from Crassostrea gigas concerning amino acid sequence similarity. Based on homologous cloning and RACE PCR, the full-length cDNA of carbonic anhydrase from Magallana hongkongensis (designated as MhCA) was cloned and sequenced. The cDNA of MhCA encodes a 315-amino-acid protein with 89.74% homology to carbonic anhydrase derived from Crassostrea gigas . Molecular docking revealed that the two zinc ions primarily form coordination bonds with histidine residues in the MhCA protein. These results strongly suggest that MhCA is a novel zinc-binding protein in Magallana hongkongensis .