Publications

• Yeast plasma membrane ATPase is essential for growth and has homology with (Na+ + K+), K+- and Ca2+-ATPases.

  Serrano R., Kielland-Brandt M.C., Fink G.R.

  The plasma membrane ATPase of plants and fungi is a hydrogen ion pump. The proton gradient generated by the enzyme drives the active transport of nutrients by H+-symport. In addition, the external acidification in plants and the internal alkalinization in fungi, both resulting from activation of t ... >> More

  The plasma membrane ATPase of plants and fungi is a hydrogen ion pump. The proton gradient generated by the enzyme drives the active transport of nutrients by H+-symport. In addition, the external acidification in plants and the internal alkalinization in fungi, both resulting from activation of the H+ pump, have been proposed to mediate growth responses. This ATPase has a relative molecular mass (Mr) similar to those of the Na+-, K+- and Ca2+-ATPases of animal cells and, like these proteins, forms an aspartylphosphate intermediate. We have cloned, mapped and sequenced the gene encoding the yeast plasma membrane ATPase (PMA1) and report here that it maps to chromosome VII adjacent to LEU1. The strong homology between the amino-acid sequence encoded by PMA1 and those of (Na+ + K+), Na+-, K+- and Ca2+-ATPases is consistent with the notion that the family of cation pumps which form a phosphorylated intermediate evolved from a common ancestral ATPase. The function of the PMA1 gene is essential because a null mutation is lethal in haploid cells. << Less

  Nature 319:689-693(1986) [PubMed] [EuropePMC]

• H+/ATP stoichiometry of proton pumps from Neurospora crassa and Escherichia coli.

  Perlin D.S., San Francisco M.J., Slayman C.W., Rosen B.P.

  A kinetic method has been used to measure the apparent stoichiometry of H+ ions translocated per ATP split by membrane-bound [H+]-ATPases. In this method, membrane vesicles are suspended in well-buffered medium, ATP is added, and a fluorescent probe of delta pH (acridine orange) is used to detect ... >> More

  A kinetic method has been used to measure the apparent stoichiometry of H+ ions translocated per ATP split by membrane-bound [H+]-ATPases. In this method, membrane vesicles are suspended in well-buffered medium, ATP is added, and a fluorescent probe of delta pH (acridine orange) is used to detect the formation of a steady-state pH gradient. At the steady state, it is assumed that proton pumping in one direction is exactly balanced by the leak of protons in the opposite direction. The pump is then rapidly turned off by the addition of an appropriate inhibitor, and the initial rate of relaxation of delta pH is used to infer the pump rate. This rate is divided by the rate of ATP hydrolysis, measured under the same condition, to give the apparent H+/ATP stoichiometry. The method has been applied to two different [H+]-ATPases, the plasma-membrane ATPase of Neurospora (a Mr = 100,000 integral membrane protein) and the ATPase of Escherichia coli (which belongs to the F0F1 group). The Neurospora ATPase displayed an apparent stoichiometry close to 1 H+/ATP (0.82-1.23), in agreement with previous estimates from electrophysiological measurements on whole cells. In contrast, the E. coli ATPase yielded an apparent stoichiometry close to 2 H+/ATP (1.90), consistent with several published values obtained by both kinetic and thermodynamic methods for bacterial, mitochondrial, and chloroplast ATPases. << Less

  Arch Biochem Biophys 248:53-61(1986) [PubMed] [EuropePMC]

  This publication is cited by 1 other entry.

• The proton-translocating ATPase of the fungal plasma membrane.

  Goffeau A., Slayman C.W.

  Biochim Biophys Acta 639:197-223(1981) [PubMed] [EuropePMC]

• Metabolic modulation of transport coupling ratio in yeast plasma membrane H(+)-ATPase.

  Venema K., Palmgren M.G.

  The plasma membrane proton pump (H(+)-ATPase) of yeast energizes solute uptake by secondary transporters and regulates cytoplasmic pH. The addition of glucose to yeast cells stimulates proton efflux mediated by the H(+)-ATPase. A > 50-fold increase in proton extrusion from yeast cells is observed ... >> More

  The plasma membrane proton pump (H(+)-ATPase) of yeast energizes solute uptake by secondary transporters and regulates cytoplasmic pH. The addition of glucose to yeast cells stimulates proton efflux mediated by the H(+)-ATPase. A > 50-fold increase in proton extrusion from yeast cells is observed in vivo, whereas the ATPase activity of purified plasma membranes is increased maximally 8-fold after glucose treatment (Serrano, R. (1983) FEBS Lett. 156, 11-14). The low capacity of yeast cells for proton extrusion in the absence of glucose can be explained by the finding that, in H(+)-ATPase isolated from glucose-starved cells, ATP hydrolysis is essentially uncoupled from proton pumping. The number of protons transported per ATP hydrolyzed is significantly increased after glucose activation. We suggest that intrinsic uncoupling is an important mechanism for regulation of pump activity. << Less

  J Biol Chem 270:19659-19667(1995) [PubMed] [EuropePMC]

• Catalytic and regulatory sites of yeast plasma membrane H(+)-ATPase studied by directed mutagenesis.

  Serrano R., Portillo F.

  More than 35 site-directed mutants of the plasma membrane H(+)-ATPase of the yeast Saccharomyces cerevisiae have been constructed and expressed to investigate the function of N- and C-termini and of conserved amino acids. Conserved motif TGES seems to form part of both the catalytic machinery for ... >> More

  More than 35 site-directed mutants of the plasma membrane H(+)-ATPase of the yeast Saccharomyces cerevisiae have been constructed and expressed to investigate the function of N- and C-termini and of conserved amino acids. Conserved motif TGES seems to form part of both the catalytic machinery for the hydrolysis of the phosphorylated intermediate and the vanadate binding site. In addition, it is involved in the coupling of ATP hydrolysis to H+ transport. The phosphorylated intermediate is also essential for this coupling, but not for ATP hydrolysis. The aspartate residues of conserved motifs DPPR, TGD and TGDGVND (the last one) seem to form part of the ATP binding site. The positive charge of the conserved motif KGAP is important for the kinase or phosphorylating activity. A conserved proline and a conserved aspartate predicted to have a transmembrane location are essential for activity. The N-terminus contains a conserved acidic region which may be involved in assembly into the plasma membrane. All the hydrophobic stretches at the C-terminus are also required for assembly. The last 11 amino acids constitute a non-essential inhibitory domain involved in regulation of the enzyme by glucose metabolism. << Less

  Biochim Biophys Acta 1018:195-199(1990) [PubMed] [EuropePMC]