title: Solubilization, purification, and characterization of (H+, K+)ATPase from hog gastric microsomes.
abstract: The (H+,K+)ATPase-enriched microsomal fraction prepared from hog gastric mucosa by sucrose density gradient centrifugation was effectively solubilized with Emulgen, with apparent preservation of the enzyme activity, and then the ATPase was highly purified by polyethylene glycol fractionation, and Blue Sepharose CL-6B and amino-hexyl Sepharose chromatographies. The purified enzyme showed a single band, with an apparent molecular mass of approximately 94 kDa, on SDS-PAGE, and exhibited both K+-ATPase and K+-stimulated-p-nitrophenyl phosphatase (pNPPase) activities. The optimum pH for the ATPase activity was 7.0. Amino acid analysis of the purified enzyme showed that it contains a large amount of hydrophobic amino acid (42%) and a small amount of glucosamine and galactosamine. The rabbit antibody monospecific for the ATPase, in the Ouchterlony double immunodiffusion and Western blotting tests, markedly inhibited both the K+-ATPase and K+-pNPPase activities.
authors.affiliation: Department of Pediatrics, Kansai Medical University, Osaka.
authors.forename: J
authors.initials: J
authors.lastname: Takaya
authors.forename: K
authors.initials: K
authors.lastname: Omori
authors.forename: S
authors.initials: S
authors.lastname: Taketani
authors.forename: Y
authors.initials: Y
authors.lastname: Kobayashi
authors.forename: Y
authors.initials: Y
authors.lastname: Tashiro
mesh_terms: D000251:Adenosine Triphosphatases / Q000302:isolation & purification*; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D005753:Gastric Mucosa / Q000201:enzymology*; D017506:H(+)-K(+)-Exchanging ATPase; D006863:Hydrogen-Ion Concentration; D007120:Immunochemistry; D008861:Microsomes / Q000201:enzymology*; D012995:Solubility; D013552:Swine
chemical_list: D000596:Amino Acids; D000251:Adenosine Triphosphatases; D017506:H(+)-K(+)-Exchanging ATPase