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Altered plasma membrane H(+)-ATPase from the Dio-9-resistant pma1-2 mutant of Schizosaccharomyces pombe.

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Ghislain, Michel [UCL] De Sadeleer, M Goffeau, André [UCL]

The pma1-2 mutation affecting the plasma membrane H(+)-ATPase of Schizosaccharomyces pombe has been selected for resistance to the antibiotic Dio-9. In membrane fractions purified from glucose-starved cells, the mutant ATPase activity is reduced by 96%, is insensitive to inhibition by vanadate and has a pH profile displaced in the acidic pH range when compared to the wild type. The maximum velocity of the H(+)-ATPase activity of plasma membranes from glucose-activated pma1-2 cells is activated 20-fold. This is in striking contrast with the wild-type ATPase activity, the maximal velocity of which is not affected by glucose. However, similar to the wild-type enzyme, glucose activation of the pma1-2 mutant H(+)-ATPase reduces the Km for MgATP 9-2 mM and shifts the optimal pH from 4.8 to 6.0-6.5. The pma1-2 mutation modifies Lys250 to a threonine, which is highly conserved in fungal and plant H(+)-ATPases. These results, compared to those reported for mutations of neighbour residues in yeast or mammalian P-type ATPases, suggest that Lys250 could play a significant role, not only in phosphate binding and/or in the E1P-E2P conformational isomerisation, but also in glucose activation of the H(+)-ATPase.

Document type Article de périodique (Journal article) – Article de recherche – Journal Article, Research Support, Non-U.S. Gov't
Publication date 1992
Language Anglais
Journal information "European journal of biochemistry / FEBS" - Vol. 209, no. 1, p. 275-9 (1992)
Peer reviewed yes
issn 0014-2956
Publication status Publié
Affiliation UCL - AGRO/CABI - Département de chimie appliquée et des bio-industries
MESH Subject Cell Membrane - enzymology ; Drug Resistance, Microbial - genetics ; Enzyme Activation - drug effects ; Glucose - pharmacology ; Glycine - chemistry ; Hydrogen-Ion Concentration ; Lysine - chemistry ; Mutation ; Organic Chemicals ; Proton-Translocating ATPases - antagonists & inhibitors - genetics - metabolism ; Schizosaccharomyces - enzymology - genetics ; Uncoupling Agents - pharmacology ; Vanadates - pharmacology
Links
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Bibliographic reference Ghislain, Michel ; De Sadeleer, M ; Goffeau, André. Altered plasma membrane H(+)-ATPase from the Dio-9-resistant pma1-2 mutant of Schizosaccharomyces pombe.. In: European journal of biochemistry / FEBS, Vol. 209, no. 1, p. 275-9 (1992)
Permanent URL http://hdl.handle.net/2078.1/9611