Subunit composition and Ca2+-ATPase activity of the vacuolar ATPase from barley roots

摘要: Abstract The vacuolar ATPase was purified from a tonoplast-enriched membrane fraction barley ( Hordeum vulgare cv CM72) roots. membranes were solubilized with Triton X-100 and the proteins separated by chromatography on Sephacryl S-400 followed fast protein liquid Mono-Q column. inhibited up to 90% KNO 3 or 80% dicyclohexylcarbodiimide (DCCI). resolved into polypeptides of 115, 68, 53, 45, 42, 34, 32, 17, 13, 12 kDa. An additional purification step centrifugation glycerol gradient did not result in loss any polypeptide bands increased specific activity ATPase. Antibodies against holoenzyme proton transport native Two peaks Ca 2+ -ATPase obtained A peak copurified during all steps NO − DCCI. It is proposed that this partial reaction plant second greatly retarded column eluted after main peak. significantly major component ATP hydrolysis membranes.