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Some properties of membrane‐bound, solubilized and reconstituted into liposomes H + ‐ATPase of vacuoles of Saccharomyces carlsbergensis
Author(s) -
Lichko L.P.,
Okorokov L.A.
Publication year - 1984
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(84)81164-3
Subject(s) - nigericin , atpase , protonophore , vacuole , biochemistry , valinomycin , chemistry , liposome , membrane , vanadate , enzyme , biophysics , biology , cytoplasm
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 NO 3 − , 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.