Immunochemical identification of exposed regions of the Na+,K+‐ATPase α‐subunit

FEBS Letters

Nagano, Yoshine¹  Matsui, Hideo¹  Homareda, Haruo¹ 

[1] Department of Biochemistry, Kyorin University School of Medicine, Mitaka, Tokyo 181, Japan

关键词: Na+;    K+-ATPase;    Antibody;    Immunoprecipitation;    cDNA;   

DOI  :  10.1016/0014-5793(93)81048-5

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

A polyclonal antibody against the Na⁺,K⁺-ATPase holoenzyme was prepared. This antibody recognized native Na⁺,K⁺-ATPase and inhibited its activity. The peptide fragments corresponding to various regions of the Na⁺,K⁺-ATPase α-subunit, which were synthesized from the cDNA, were immunoprecipitated with the antibody, and the M32-D75, M158-D197 and M470-V552 fragments (the latter included K508, a putative ATP binding site) were identified as the epitopes. The M267-I442 fragment, which included a phosphorylation site at D376, and the C-terminal one-third of the α-subunit from M615 to the C-terminus, were not detected using this antibody. These results suggest that at least three regions on the Na⁺,K⁺-ATPase α-subunit, M32-D75, M158-D197 and M470-V552, cover its exposed regions, and that some of them are essential for ATPase activity.

【 授权许可】

Unknown