[38] Plasma membrane ATPase from the yeast Schizosaccharomyces pombe

Dufour, Jean‐Pierre; Amory, Antoine; Goffeau, André

doi:10.1016/0076-6879(88)57100-8

Cited by 71 publications

(27 citation statements)

References 18 publications

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“…Isolated plasma membranes (0.1 or 0.2 g per well) were incubated with 4 mM ATP, 5 mM MgCl 2 in 300 mM Tris-glycine buffer (pH 9.0) in a final volume of 25 l. To reduce background, 0.2 mM ammonium molybdate, 50 mM KNO 3 , and 10 mM NaN 3 , respectively, were added (9,19). In a control reaction, OM (20 g/ml) was added to the assay to determine the OM-sensitive ATPase activity.…”

Section: Methodsmentioning

confidence: 99%

Generating Symmetry in the Asymmetric ATP-binding Cassette (ABC) Transporter Pdr5 from Saccharomyces cerevisiae

Gupta

Kueppers

Hanekop

et al. 2014

Journal of Biological Chemistry

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Background: Fungal multidrug efflux pumps possess a degenerate nucleotide-binding site (NBS). Results: Restoring all the nonconserved amino acids in the degenerate NBS of Pdr5 leads to complete loss of function of the protein. Conclusion:The degenerate NBS is essential and acts as a structural platform supporting the canonical NBS. Significance: This is the first study dealing with the entire degenerate NBS and its functional role.

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Section: Methodsmentioning

confidence: 99%

Generating Symmetry in the Asymmetric ATP-binding Cassette (ABC) Transporter Pdr5 from Saccharomyces cerevisiae

Gupta

Kueppers

Hanekop

et al. 2014

Journal of Biological Chemistry

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“…For determination of the apparent K, for MgATP and maximal velocity V, the ATP concentrations were varying over 0.15 -16 mM, with an excess of 2 mM MgS04 over ATP. After 4 -30 min, the reaction was stopped by addition of 0.3 ml and 6% SDS; inorganic phosphate was measured as described [18], cxcept that 0.25 ml ammonium molybdate and 0.25 ml ELON developing reagent (Kodak) were added. The rate of hydrolysis was linear with the assay time.…”

Section: Atpase Assaysmentioning

confidence: 99%

Altered plasma membrane H⁺‐ATPase from the Dio‐9‐resistant pmal‐2 mutant of Schizosaccharomyces pombe

Ghislain

Sadeleer

Goffeau

1992

European Journal of Biochemistry

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The pmul-2 mutation affecting the plasma membrane H + -ATPase of Schizosacchuromycespombe has been selected for resistance to the antibiotic Dio-9. In membrane fractions purified from glucosestarved 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 pmul-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 thepmal-2 mutant Hf-ATPase reduces the K , for MgATP 9 -2 mM and shifts the optimal pH from The pmal-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 EIP-E2P conformational isomerisation, but also in glucose activation of the HC-ATPase. [7] and Candida albicans [S]. The closely related polypeptides are folded similarly in the plasma membrane, with 8 -12 putative transmembrane segments delimiting two major cytoplasmic regions. The largest hydrophilic region contains the nucleotide-binding and phosphorylation domains [9]. A smaller hydrophilic domain, rcferred to as an energy-transduction or P-stranded [lo] domain, is believed to be essential for protein phosphatase activity [9] and/or for conformational changes at the level of the phosphoenzyme [I 1 I. The pmal-1 mutation, affecting the fission yeast H +-ATPase, was originally selected on the basis of Dio-9 resistance [12] and was later shown to result from rcplacemcnt of Gly268 by an aspartate [7]. The ATPase activity is decreased and highly resistant to vanadate [13]. The kinetic properties of the mutant enzyme are opposite to changes resulting from activation of the S. cerevisiae H ' -ATPase by glucose [14]. It has therefore been suggested that the pmal-1 mutation modifies the physiological regulation of ATPase activity by glucose [15].In this paper, we report on the properties of the novel Dio-9-rcsistant mutation pmal-2 which affects the residue Lys250Correspondence to A. Goffeau, Unite de Biochimie Physiologiquc, Place Croix du Sud 2/20, B-I 348 Louvain-la-Neuvc, BelgiumEnzyme. H+-ATPase, ATP phosphohydrolase (EC 3.6.1.35).located close to Gly268 within the small hydrophilic domain. We also compare the glucose activation of the pmal-1 and pmaf-2 mutant H+-ATPases. MATERIALS AND METHODS Yeast strains and mediaThepmal-1 strain (JV66) andpmal-2 strains (JV27, JV28, JV29 and JV37) are derivatives of the wild-type strain 97211-ade7-413. The mutants were isolated after mutagenesis with Nmethyl-N'-nitro-N-nitrosoguanidine and selection for growth resistance to 131. Yeast strains were grown in 5.8% (massivol.) glucose, 2% (ma...

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“…For sarcolemmal Caz+ ATPase activity, the rate of ATP hydrolysis was determined at 37°C (pH 7.4) by measuring the amount of P, liberated during a 15-min incubation of the vesicles (40 pg/ml) using the same colorimetric method [33]. This was done at pCa 6.0 for 1 -(3,4-dimethoxyphenyI)-3-dodecanone and pCa,, (6.46) for ellagic acid in order to be able to see a potential activation for each compound.…”

Section: Myocyte Isolationmentioning

confidence: 99%

“…The rate of ATP hydrolysis was determined at 30°C by measuring the amount of P, liberated during a 15-min incubation of the SR vesicles (40 pg/ml and 6 pg/ml for cardiac and skeletal muscle, respectively) using a colorimetric method [33]. The medium containing (in mmol/l) 100 KCI, 2 MgCl,, 50 Mops/KOH, pH 6.8, 2 Na'ATP, 1 EGTA and different CaCl, concentrations to yield the desired [Ca2+lfree that were calculated using Fabiato's computer programme [3 11.…”

Section: Myocyte Isolationmentioning

confidence: 99%

Mechanism of Action of Sarcoplasmic Reticulum Calcium‐Uptake Activators

Berrebi-Bertrand¹,

Lahouratate²,

Lahouratate³

et al. 1997

European Journal of Biochemistry

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The Ca'+ uptake by the sarcoplasmic reticulum (SR) can be affected by direct modulation of the Ca2+ pump or by removing the inhibitory effect of dephosphorylated phospholamban. The effect of these mechanisms was assessed using ellagic acid and 1-(3,4-dimethoxypheny1)-3-dodecanone. Both compounds (30 pmolll) enhanced SR-Ca" uptake in rabbit cardiomyocytes by 65.3 t 13 % and 44.3 5 6.7 % for 1-(3,4-dimethoxyphenyl)-3-dodecanone and ellagic acid, respectively (at pCa 6.2). A similar effect was observed in cardiac SR microsomes (59.5 t 7.4% and 45.1 t 6.7) with 30 pmol/l 1-(3,4-dimethodoxyphenyl)-3-dodecanone and ellagic acid, respectively. 1-(3,4-DimethoxyphenyI)-3-dodecanone increased Ca" storage by cardiac SR microsomes mainly at high [Ca'+] with a 57% increase of V,,,,,, whereas ellagic acid increased V,,, to a smaller extent (22%) and stimulated Ca2+ uptake at lower [CaZ+] with a leftward-shift of the pCdATPase relationship by pCa 0.24. Ellagic acid also differed from 1-(3,4-dimethoxylphenyl)-3-dodecanone in that it produced a Ca" sensitizing effect only in cardiac SR microsomes (by pCa 0.3) whereas 1-(3,4-dimethoxyphenyl)-3-dodecanone stimulated the ATPase, at saturating Ca2+, in both cardiac and skeletal muscle SR vesicles. It is suggested that 1-(3.4-dimethoxypheny1)-3-dodecanone stimulates directly the Ca2+-ATPase activity, in contrast to ellagic acid which enhances the cardiac SR-CaZ+ uptake by interacting with phospholamban, as confirmed by the lack of additive effect between ellagic acid and monoclonal antibodies raised against phospholamban. 1-(3,4-dimethoxyphenyl)-3-dodecanone and ellagic acid constitute attractive pharmacological tools to investigate the functional consequences of enhancing SR Ca'+, uptake by affecting different mechanisms.

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[38] Plasma membrane ATPase from the yeast Schizosaccharomyces pombe

Cited by 71 publications

References 18 publications

Generating Symmetry in the Asymmetric ATP-binding Cassette (ABC) Transporter Pdr5 from Saccharomyces cerevisiae

Generating Symmetry in the Asymmetric ATP-binding Cassette (ABC) Transporter Pdr5 from Saccharomyces cerevisiae

Altered plasma membrane H⁺‐ATPase from the Dio‐9‐resistant pmal‐2 mutant of Schizosaccharomyces pombe

Mechanism of Action of Sarcoplasmic Reticulum Calcium‐Uptake Activators

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[38] Plasma membrane ATPase from the yeast Schizosaccharomyces pombe

Cited by 71 publications

References 18 publications

Generating Symmetry in the Asymmetric ATP-binding Cassette (ABC) Transporter Pdr5 from Saccharomyces cerevisiae

Generating Symmetry in the Asymmetric ATP-binding Cassette (ABC) Transporter Pdr5 from Saccharomyces cerevisiae

Altered plasma membrane H+‐ATPase from the Dio‐9‐resistant pmal‐2 mutant of Schizosaccharomyces pombe

Mechanism of Action of Sarcoplasmic Reticulum Calcium‐Uptake Activators

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Altered plasma membrane H⁺‐ATPase from the Dio‐9‐resistant pmal‐2 mutant of Schizosaccharomyces pombe