title: Two-dimensional gel electrophoresis and FTIR spectroscopy reveal both forms of yeast plasma membrane H(+)-ATPase in activated and basal-level enzyme preparations.
abstract: Plasma membrane H(+)-ATPase of the yeast Saccharomyces cerevisiae was isolated and purified in its two forms, the activated A-ATPase from glucose-metabolizing cells, and the basal-level B-ATPase from cells with endogenous metabolism only. Using two-dimensional gel electrophoretic analysis, we showed that both enzyme preparations are actually mixtures of the non-active, i.e. non-phosphorylated, and the active, i.e. phosphorylated, forms of the enzyme. Previous deliberations suggesting that the B-ATPase displays some activity which is lower than that of A-ATPase were apparently wrong. It seems that, molecularly speaking, the B-form is actually not active at all, and what activity we measure in our preparation is due to an admixture of the true active form (A-form). Fourier transform infrared spectroscopic study of the secondary structure and particularly thermal denaturation data suggest the possibility that the two enzyme forms interact to form complexes less stable than the single forms. On the whole then, there apparently is a different ratio of the active and inactive forms and/or complexes between the two forms present in all enzyme preparations.
authors.affiliation: Institute of Animal Physiology and Genetics, Czech Academy of Sciences, Libechov, Czech Republic. lapathitis@iapg.cas.cz
authors.forename: G
authors.initials: G
authors.lastname: Lapathitis
authors.forename: F
authors.initials: F
authors.lastname: Tanfani
authors.forename: A
authors.initials: A
authors.lastname: Kotyk
authors.forename: E
authors.initials: E
authors.lastname: Bertoli
mesh_terms: D002462:Cell Membrane / Q000201:enzymology*; D015180:Electrophoresis, Gel, Two-Dimensional; D004789:Enzyme Activation; D010766:Phosphorylation; D017433:Protein Structure, Secondary; D006180:Proton-Translocating ATPases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D017550:Spectroscopy, Fourier Transform Infrared; D015003:Yeasts / Q000201:enzymology*
chemical_list: D006180:Proton-Translocating ATPases