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Immunochemical identification of exposed regions of the Na + ,K + ‐ATPase α‐subunit
Author(s) -
Homareda Haruo,
Nagano Yoshine,
Matsui Hideo
Publication year - 1993
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(93)81048-5
Subject(s) - polyclonal antibodies , microbiology and biotechnology , protein subunit , atpase , antibody , chemistry , epitope , g alpha subunit , peptide , biochemistry , biology , enzyme , immunology , gene
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.