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

Updated：2023-05-16

- Na+/K+-ATPase α-subunit in swimming crab Portunus trituberculatus: molecular cloning, characterization, and expression under low salinity stress
- Journal of Oceanology and Limnology Vol. 33, Issue 4, Pages: 828-837(2015)
- 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：
- DOI：10.1007/s00343-015-4018-9
  CLC：
- Online First：18 March 2015，
  
  Published：2015
- Accepted：
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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：

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.

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references

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