Mutational study on the roles of disulfide bonds in the beta-subunit of gastric H+,K+-ATPase

The Journal of Biological Chemistry
Tohru KimuraShinji Asano

Abstract

The gastric proton pump, H(+),K(+)-ATPase, consists of the catalytic alpha-subunit and the non-catalytic beta-subunit. Correct assembly between the alpha- and beta-subunits is essential for the functional expression of H(+),K(+)-ATPase. The beta-subunit contains nine conserved cysteine residues; two are in the cytoplasmic domain, one in the transmembrane domain, and six in the ectodomain. The six cysteine residues in the ectodomain form three disulfide bonds. In this study, we replaced each of the cysteine residues of the beta-subunit with serine individually and in several combinations. The mutant beta-subunits were co-expressed with the alpha-subunit in human embryonic kidney 293 cells, and the role of each cysteine residue or disulfide bond in the alpha/beta assembly, stability, and cell surface delivery of the alpha- and beta-subunits and H(+),K(+)-ATPase activity was studied. Mutant beta-subunits with a replacement of the cytoplasmic and transmembrane cysteines preserved H(+),K(+)-ATPase activity. All the mutant beta-subunits with replacement(s) of the extracellular cysteines did not assemble with the alpha-subunit, resulting in loss of H(+),K(+)-ATPase activity. These mutants did not permit delivery of the alpha-subunit t...Continue Reading

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Citations

Feb 27, 2008·Journal of Clinical Biochemistry and Nutrition·Masahiko NakamuraKanji Tsuchimoto
May 24, 2003·Annals of the New York Academy of Sciences·Shinji AsanoNoriaki Takeguchi
Jan 8, 2008·The Journal of Biological Chemistry·Takuto FujiiHideki Sakai
Jan 28, 2004·Biological & Pharmaceutical Bulletin·Shinji AsanoNoriaki Takeguchi
Nov 2, 2019·Physiological Reviews·Amy C EngevikJames R Goldenring
May 30, 2003·American Journal of Physiology. Cell Physiology·O VaginG Sachs
Jun 27, 2003·The Journal of Biological Chemistry·Melissa D LaugheryJack H Kaplan
Dec 31, 2003·The Journal of Biological Chemistry·Shinji AsanoMagotoshi Morii
Jun 11, 2019·Nihon yakurigaku zasshi. Folia pharmacologica Japonica·Takuto FujiiHideki Sakai
Jan 17, 2014·Biology Open·Yuji TakahashiHideki Sakai

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