Na+/K+-ATPase α-subunit in swimming crab Portunus trituberculatus: molecular cloning, characterization, and expression under low salinity stress

Xiaolin Han; Ping Liu; Baoquan Gao; Haofeng Wang; Yafei Duan; Wenfei Xu; Ping Chen

doi:10.1007/s00343-015-4018-9

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Na+/K+-ATPase α-subunit in swimming crab Portunus trituberculatus: molecular cloning, characterization, and expression under low salinity stress

OriginalPaper | Updated：2023-05-16

- Na+/K+-ATPase α-subunit in swimming crab Portunus trituberculatus: molecular cloning, characterization, and expression under low salinity stress
- Na+/K+-ATPase α-subunit in swimming crab Portunus trituberculatus: molecular cloning, characterization, and expression under low salinity stress
- 海洋湖沼学报（英文） 2015年33卷第4期页码：828-837
- Affiliations：
  
  1. Key Laboratory of Sustainable Development of Marine Fisheries Ministry of Agriculture Yellow Sea Fisheries Research Institute Chinese Academy of Fishery Sciences
  2. Dalian Ocean University
- Author bio：
- Funds：
  
  Supported by the National High Technology Research and Development Program of China (863 Program) (No. 2012AA10A409), the National Natural Science Foundation of China (No. 41306177), and the Special Scientific Research Funds for Central Non-Profit Institutes, Yellow Sea Fisheries Research Institutes (No. 20603022013027)
- DOI：10.1007/s00343-015-4018-9
  中图分类号：
- 网络首发：2015-03-18，
  
  纸质出版：2015
- Accepted：
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Na+/K+-ATPase α-subunit in swimming crab Portunus trituberculatus: molecular cloning, characterization, and expression under low salinity stress[J]. 海洋湖沼学报（英文）, 2015,33(4):828-837.

Han, X., Liu, P., Gao, B. et al. Na+/K+-ATPase α-subunit in swimming crab Portunus trituberculatus: molecular cloning, characterization, and expression under low salinity stress., Chin. J. Ocean. Limnol. 33, 828–837 (2015). https://doi.org/10.1007/s00343-015-4018-9
Na+/K+-ATPase α-subunit in swimming crab Portunus trituberculatus: molecular cloning, characterization, and expression under low salinity stress[J]. 海洋湖沼学报（英文）, 2015,33(4):828-837. DOI： 10.1007/s00343-015-4018-9.

Han, X., Liu, P., Gao, B. et al. Na+/K+-ATPase α-subunit in swimming crab Portunus trituberculatus: molecular cloning, characterization, and expression under low salinity stress., Chin. J. Ocean. Limnol. 33, 828–837 (2015). https://doi.org/10.1007/s00343-015-4018-9 DOI：

摘要

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

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.

关键词

Keywords

references

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