Some properties of membrane‐bound, solubilized and reconstituted into liposomes H+‐ATPase of vacuoles of Saccharomyces carlsbergensis

FEBS Letters

Lichko, L.P.¹  Okorokov, L.A.¹ 

[1] Institute of Biochemistry and Physiology of Microorganisms, USSR Academy of Sciences, Pushchino, Moscow Region 142292, USSR

关键词: ATPase;    GTPase;    Solubilization;    Purification;    Reconstitution;    Liposome;    H+ transport;   

DOI  :  10.1016/0014-5793(84)81164-3

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Vacuoles of yeast grown in peptone medium possessed high ATPase activity (up to 1 μmol·mg protein-1·min⁻¹). Membrane-bound and solubilized ATPase activities were insensitive to vanadate and azide, but were inhibited by NO₃ ⁻, K⁺ and cyclic AMP stimulated both membrane-bound and solubilized ATPase activities. Dio-9 activated the membrane form of vacuolar ATPase 1.5-2-fold and did not affect the solubilized enzyme. Solubilized and partially purified vacuolar ATPase was reconstituted with soy-bean phospholipids by a freeze-thaw procedure. ATPase activities in native vacuoles and proteoliposomes were stimulated effectively by Dio-9, the protonophore FCCP and ionophores valinomycin and nigericin. ATP-dependent H⁺ transport into proteoliposomes was also shown by quenching of ACMA fluorescence. Vacuolar and partially purified ATPase preparations possessed also GTPase activity. Unlike ATPase, however, GTPase was not incorporated as a proton pump into liposomes.

【 授权许可】

Unknown