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J Biol Chem
2010 Dec 10;28550:39366-79. doi: 10.1074/jbc.M110.133470.
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Deceleration of the E1P-E2P transition and ion transport by mutation of potentially salt bridge-forming residues Lys-791 and Glu-820 in gastric H+/K+-ATPase.
Dürr KL
,
Seuffert I
,
Friedrich T
.
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A lysine residue within the highly conserved center of the fifth transmembrane segment in P(IIC)-type ATPase α-subunits is uniquely found in H,K-ATPases instead of a serine in all Na,K-ATPase isoforms. Because previous studies suggested a prominent role of this residue in determining the electrogenicity of non-gastric H,K-ATPase and in pK(a) modulation of the proton-translocating residues in the gastric H,K-ATPases as well, we investigated its functional significance for ion transport by expressing several Lys-791 variants of the gastric H,K-ATPase in Xenopus oocytes. Although the mutant proteins were all detected at the cell surface, none of the investigated mutants displayed any measurable K(+)-induced stationary currents. In Rb(+) uptake measurements, replacement of Lys-791 by Arg, Ala, Ser, and Glu substantially impaired transport activity and reduced the sensitivity toward the E(2)-specific inhibitor SCH28080. Furthermore, voltage clamp fluorometry using a reporter site in the TM5/TM6 loop for labeling with tetra-methylrhodamine-6-maleimide revealed markedly changed fluorescence signals. All four investigated mutants exhibited a strong shift toward the E(1)P state, in agreement with their reduced SCH28080 sensitivity, and an about 5-10-fold decreased forward rate constant of the E(1)P ↔ E(2)P conformational transition, thus explaining the E(1)P shift and the reduced Rb(+) transport activity. When Glu-820 in TM6 adjacent to Lys-791 was replaced by non-charged or positively charged amino acids, severe effects on fluorescence signals and Rb(+) transport were also observed, whereas substitution by aspartate was less disturbing. These results suggest that formation of an E(2)P-stabilizing interhelical salt bridge is essential to prevent futile proton exchange cycles of H(+) pumping P-type ATPases.
Abramson,
Structure and mechanism of the lactose permease of Escherichia coli.
2003, Pubmed
Abramson,
Structure and mechanism of the lactose permease of Escherichia coli.
2003,
Pubmed
Anderson,
pH-induced denaturation of proteins: a single salt bridge contributes 3-5 kcal/mol to the free energy of folding of T4 lysozyme.
1990,
Pubmed
Arnold,
The SWISS-MODEL workspace: a web-based environment for protein structure homology modelling.
2006,
Pubmed
Asano,
Mutational analysis of putative SCH 28080 binding sites of the gastric H+,K+-ATPase.
1997,
Pubmed
Axelsen,
Evolution of substrate specificities in the P-type ATPase superfamily.
1998,
Pubmed
Bordoli,
Protein structure homology modeling using SWISS-MODEL workspace.
2009,
Pubmed
Buffin-Meyer,
K depletion modifies the properties of Sch-28080-sensitive K-ATPase in rat collecting duct.
1997,
Pubmed
Burnay,
Bufo marinus bladder H-K-ATPase carries out electroneutral ion transport.
2001,
Pubmed
,
Xenbase
Burnay,
Electrogenicity of Na,K- and H,K-ATPase activity and presence of a positively charged amino acid in the fifth transmembrane segment.
2003,
Pubmed
,
Xenbase
Chiesi,
ATP synthesis catalyzed by the purified erythrocyte Ca-ATPase in the absence of calcium gradients.
1984,
Pubmed
Chmouliovsky,
Increase in ATP by reversal of the Na-K-pump in mammalian non-myelinated nerve fibres.
1974,
Pubmed
Cougnon,
The rat distal colon P-ATPase alpha subunit encodes a ouabain-sensitive H+, K+-ATPase.
1996,
Pubmed
,
Xenbase
Cougnon,
Does the colonic H,K-ATPase also act as an Na,K-ATPase?
1998,
Pubmed
,
Xenbase
De Pont,
The E1/E2-preference of gastric H,K-ATPase mutants.
2003,
Pubmed
De Weer,
Current generated by backward-running electrogenic Na pump in squid giant axons.
,
Pubmed
Dunten,
Role of the charge pair aspartic acid-237-lysine-358 in the lactose permease of Escherichia coli.
1993,
Pubmed
Dürr,
Characterization of Na,K-ATPase and H,K-ATPase enzymes with glycosylation-deficient beta-subunit variants by voltage-clamp fluorometry in Xenopus oocytes.
2008,
Pubmed
,
Xenbase
Dürr,
Functional significance of E2 state stabilization by specific alpha/beta-subunit interactions of Na,K- and H,K-ATPase.
2009,
Pubmed
,
Xenbase
Dürr,
E2P state stabilization by the N-terminal tail of the H,K-ATPase beta-subunit is critical for efficient proton pumping under in vivo conditions.
2009,
Pubmed
,
Xenbase
Efthymiadis,
Conditions for a backward-running Na+/K+ pump in Xenopus oocytes.
1991,
Pubmed
,
Xenbase
Garrahan,
Driving the sodium pump backwards to form adenosine triphosphate.
1966,
Pubmed
Garrahan,
The incorporation of inorganic phosphate into adenosine triphosphate by reversal of the sodium pump.
1967,
Pubmed
Geering,
The functional role of beta subunits in oligomeric P-type ATPases.
2001,
Pubmed
Geibel,
Conformational dynamics of the Na+/K+-ATPase probed by voltage clamp fluorometry.
2003,
Pubmed
,
Xenbase
Geibel,
Conformational dynamics of Na+/K+- and H+/K+-ATPase probed by voltage clamp fluorometry.
2003,
Pubmed
,
Xenbase
Gottardi,
Molecular requirements for the cell-surface expression of multisubunit ion-transporting ATPases. Identification of protein domains that participate in Na,K-ATPase and H,K-ATPase subunit assembly.
1993,
Pubmed
Gupta,
Evidence for a salt bridge between transmembrane segments 5 and 6 of the yeast plasma-membrane H+-ATPase.
1998,
Pubmed
Helmich-de Jong,
Direct evidence for an ADP-sensitive phosphointermediate of (K+ + H+)-ATPase.
1985,
Pubmed
Hermsen,
Mimicking of K+ activation by double mutation of glutamate 795 and glutamate 820 of gastric H+,K+-ATPase.
2001,
Pubmed
Hermsen,
Mutagenesis of glutamate 820 of the gastric H+,K+-ATPase alpha-subunit to aspartate decreases the apparent ATP affinity.
1999,
Pubmed
Humphrey,
VMD: visual molecular dynamics.
1996,
Pubmed
Jaisser,
Mechanisms of urinary K+ and H+ excretion: primary structure and functional expression of a novel H,K-ATPase.
1993,
Pubmed
,
Xenbase
King,
The interaction between aspartic acid 237 and lysine 358 in the lactose carrier of Escherichia coli.
1991,
Pubmed
Knowles,
Formation of adenosine triphosphate from Pi and adenosine diphosphate by purified Ca-2+-adenosine triphosphatase.
1975,
Pubmed
Koenderink,
A conformation-specific interhelical salt bridge in the K+ binding site of gastric H,K-ATPase.
2004,
Pubmed
Lew,
Net synthesis of ATP by reversal of the sodium pump.
1970,
Pubmed
Lorentzon,
Inhibitory effects of cations on the gastric H+, K+ -ATPase. A potential-sensitive step in the K+ limb of the pump cycle.
1988,
Pubmed
Lorenz,
Heteromultimeric CLC chloride channels with novel properties.
1996,
Pubmed
,
Xenbase
Mendlein,
Interaction of a K(+)-competitive inhibitor, a substituted imidazo[1,2a] pyridine, with the phospho- and dephosphoenzyme forms of H+, K(+)-ATPase.
1990,
Pubmed
Morth,
Crystal structure of the sodium-potassium pump.
2007,
Pubmed
Munson,
Inhibitor and ion binding sites on the gastric H,K-ATPase.
2005,
Pubmed
Nissen,
One way for the gastric proton pump.
2009,
Pubmed
Pedersen,
Crystal structure of the plasma membrane proton pump.
2007,
Pubmed
Rajendran,
Ouabain-sensitive H,K-ATPase functions as Na,K-ATPase in apical membranes of rat distal colon.
2000,
Pubmed
Sachs,
A nonelectrogenic H+ pump in plasma membranes of hog stomach.
1976,
Pubmed
Sahin-Tóth,
Functional interactions between putative intramembrane charged residues in the lactose permease of Escherichia coli.
1992,
Pubmed
Schuurmans Stekhoven,
Free protons do not substitute for sodium ions in buffer-mediated phosphorylation of (Na+ + K+)-ATPase.
1986,
Pubmed
Shin,
The gastric HK-ATPase: structure, function, and inhibition.
2009,
Pubmed
Stengelin,
Kinetics of transient pump currents generated by the (H,K)-ATPase after an ATP concentration jump.
1993,
Pubmed
Sun,
Contributions of engineered surface salt bridges to the stability of T4 lysozyme determined by directed mutagenesis.
1991,
Pubmed
Swarts,
K(+)-independent gastric H(+),K(+)-atpase activity. Dissociation of K(+)-independent dephosphorylation and preference for the E1 conformation by combined mutagenesis of transmembrane glutamate residues.
2001,
Pubmed
Swarts,
Constitutive activation of gastric H+,K+-ATPase by a single mutation.
1998,
Pubmed
Swarts,
Conformation-dependent inhibition of gastric H+,K+-ATPase by SCH 28080 demonstrated by mutagenesis of glutamic acid 820.
1999,
Pubmed
Taniguchi,
Synthesis of adenosine triphosphate and exchange between inorganic phosphate and adenosine triphosphate in sodium and potassium ion transport adenosine triphosphatase.
1975,
Pubmed
Thomas,
Membrane current and intracellular sodium changes in a snail neurone during extrusion of injected sodium.
1969,
Pubmed
Vagin,
Mutational analysis of the K+-competitive inhibitor site of gastric H,K-ATPase.
2001,
Pubmed
Wallmark,
Inhibition of gastric (H+ + K+)-ATPase by the substituted benzimidazole, picoprazole.
1983,
Pubmed
Wüthrich,
Net ATP synthesis by running the red cell calcium pump backwards.
1979,
Pubmed
de Meis,
A new mechanism by which an H+ concentration gradient drives the synthesis of adenosine triphosphate, pH jump, and adenosine triphosphate synthesis by the Ca2+-dependnet adenosine triphosphatase of sarcoplasmic reticulum.
1977,
Pubmed
van der Hijden,
Demonstration of the electrogenicity of proton translocation during the phosphorylation step in gastric H+K(+)-ATPase.
1990,
Pubmed
