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| Content Provider | Springer Nature Link |
|---|---|
| Author | Han, Xiaolin Liu, Ping Gao, Baoquan Wang, Haofeng Duan, Yafei Xu, Wenfei Chen, Ping |
| Copyright Year | 2015 |
| Abstract | Na+/K+-ATPases are membrane-associated enzymes responsible for the active transport of Na+ and K+ ions across cell membranes, generating chemical and electrical gradients. These enzymes’ α-subunit provides catalytic function, binding and hydrolyzing ATP, and itself becoming phosphorylated during the transport cycle. In this study, Na+/K+-ATPase α-subunit cDNA was cloned from gill tissue of the swimming crab Portunus trituberculatus by reverse-transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA end methods. Analysis of the nucleotide sequence revealed that the cDNA had a full-length of 3 833 base pairs (bp), with an open reading frame of 3 120 bp, 5′ untranslated region (UTR) of 317 bp, and 3′ UTR of 396 bp. The sequence encoded a 1 039 amino acid protein with a predicted molecular weight of 115.57 kDa and with estimated pI of 5.21. It was predicted here to possess all expected features of Na+/K+-ATPase members, including eight transmembrane domains, putative ATP-binding site, and phosphorylation site. Comparison of amino acid sequences showed that the P. trituberculatus α-subunit possessed an overall identity of 75%–99% to that of other organisms. Phylogenetic analysis revealed that this α-subunit was in the same category as those of crustaceans. Quantitative real-time RT-PCR analysis indicated that this α-subunit’s transcript were most highly expressed in gill and lowest in muscle. RT-PCR analysis also revealed that α-subunit expression in crab gill decreased after 2 and 6 h, but increased after 12, 24, 48, and 72 h. In addition, α-subunit expression in hepatopancreas of crab decreased after 2-72 h. These facts indicated that the crab’s Na+/K+-ATPase α-subunit was potentially involved in the observed acute response to low salinity stress. |
| Starting Page | 828 |
| Ending Page | 837 |
| Page Count | 10 |
| File Format | |
| ISSN | 02544059 |
| Journal | Chinese Journal of Oceanology and Limnology |
| Volume Number | 33 |
| Issue Number | 4 |
| e-ISSN | 19935005 |
| Language | English |
| Publisher | Science Press |
| Publisher Date | 2015-03-18 |
| Publisher Place | Heidelberg |
| Access Restriction | Subscribed |
| Subject Keyword | cloning expression Na+/K+-ATPase α-subunit Portunus trituberculatus salinity Oceanography |
| Content Type | Text |
| Resource Type | Article |
| Subject | Water Science and Technology Oceanography |
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