Purification, Characterization, cDNA Cloning, and Bioinformatic Analysis of Zinc-Binding Protein from <i>Magallana hongkongensis</i>
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 <i>Magallana hongkongensis</i> using two kinds of gel filtration chromatograms. Sodium dodecyl sulfat...
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MDPI AG
2024-02-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/29/4/900 |
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| author | Citing Chen Wan Li Jialong Gao Wenhong Cao Xiaoming Qin Huina Zheng Haisheng Lin Zhongqin Chen |
| author_facet | Citing Chen Wan Li Jialong Gao Wenhong Cao Xiaoming Qin Huina Zheng Haisheng Lin Zhongqin Chen |
| author_sort | Citing Chen |
| collection | DOAJ |
| description | 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 <i>Magallana hongkongensis</i> 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 <i>Crassostrea gigas</i> concerning amino acid sequence similarity. Based on homologous cloning and RACE PCR, the full-length cDNA of carbonic anhydrase from <i>Magallana hongkongensis</i> (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 <i>Crassostrea gigas</i>. 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 <i>Magallana hongkongensis</i>. |
| first_indexed | 2024-03-07T22:19:30Z |
| format | Article |
| id | doaj.art-0bcea077420049a4b6415e2dd3dc464b |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-07T22:19:30Z |
| publishDate | 2024-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-0bcea077420049a4b6415e2dd3dc464b2024-02-23T15:29:11ZengMDPI AGMolecules1420-30492024-02-0129490010.3390/molecules29040900Purification, Characterization, cDNA Cloning, and Bioinformatic Analysis of Zinc-Binding Protein from <i>Magallana hongkongensis</i>Citing Chen0Wan Li1Jialong Gao2Wenhong Cao3Xiaoming Qin4Huina Zheng5Haisheng Lin6Zhongqin Chen7College of Food Science and Technology, Guangdong Ocean University, Zhanjiang 524088, ChinaCollege of Food Science and Technology, Guangdong Ocean University, Zhanjiang 524088, ChinaCollege of Food Science and Technology, Guangdong Ocean University, Zhanjiang 524088, ChinaCollege of Food Science and Technology, Guangdong Ocean University, Zhanjiang 524088, ChinaCollege of Food Science and Technology, Guangdong Ocean University, Zhanjiang 524088, ChinaCollege of Food Science and Technology, Guangdong Ocean University, Zhanjiang 524088, ChinaCollege of Food Science and Technology, Guangdong Ocean University, Zhanjiang 524088, ChinaCollege of Food Science and Technology, Guangdong Ocean University, Zhanjiang 524088, ChinaOysters 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 <i>Magallana hongkongensis</i> 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 <i>Crassostrea gigas</i> concerning amino acid sequence similarity. Based on homologous cloning and RACE PCR, the full-length cDNA of carbonic anhydrase from <i>Magallana hongkongensis</i> (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 <i>Crassostrea gigas</i>. 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 <i>Magallana hongkongensis</i>.https://www.mdpi.com/1420-3049/29/4/900<i>Magallana hongkongensis</i>zinc-binding proteincDNA cloningcarbonic anhydrase |
| spellingShingle | Citing Chen Wan Li Jialong Gao Wenhong Cao Xiaoming Qin Huina Zheng Haisheng Lin Zhongqin Chen Purification, Characterization, cDNA Cloning, and Bioinformatic Analysis of Zinc-Binding Protein from <i>Magallana hongkongensis</i> Molecules <i>Magallana hongkongensis</i> zinc-binding protein cDNA cloning carbonic anhydrase |
| title | Purification, Characterization, cDNA Cloning, and Bioinformatic Analysis of Zinc-Binding Protein from <i>Magallana hongkongensis</i> |
| title_full | Purification, Characterization, cDNA Cloning, and Bioinformatic Analysis of Zinc-Binding Protein from <i>Magallana hongkongensis</i> |
| title_fullStr | Purification, Characterization, cDNA Cloning, and Bioinformatic Analysis of Zinc-Binding Protein from <i>Magallana hongkongensis</i> |
| title_full_unstemmed | Purification, Characterization, cDNA Cloning, and Bioinformatic Analysis of Zinc-Binding Protein from <i>Magallana hongkongensis</i> |
| title_short | Purification, Characterization, cDNA Cloning, and Bioinformatic Analysis of Zinc-Binding Protein from <i>Magallana hongkongensis</i> |
| title_sort | purification characterization cdna cloning and bioinformatic analysis of zinc binding protein from i magallana hongkongensis i |
| topic | <i>Magallana hongkongensis</i> zinc-binding protein cDNA cloning carbonic anhydrase |
| url | https://www.mdpi.com/1420-3049/29/4/900 |
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