Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/67697
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dc.contributor.authorHsiao, Y.Y.en_US
dc.contributor.authorPan, Y.J.en_US
dc.contributor.authorHsu, S.H.en_US
dc.contributor.authorHuang, Y.T.en_US
dc.contributor.authorLiu, T.H.en_US
dc.contributor.authorLee, C.H.en_US
dc.contributor.authorLiu, P.F.en_US
dc.contributor.authorChang, W.C.en_US
dc.contributor.authorWang, Y.K.en_US
dc.contributor.authorChien, L.F.en_US
dc.contributor.authorPan, R.L.en_US
dc.date2007zh_TW
dc.date.accessioned2014-06-11T05:55:32Z-
dc.date.available2014-06-11T05:55:32Z-
dc.identifier.issn0005-2728zh_TW
dc.identifier.urihttp://hdl.handle.net/11455/67697-
dc.description.abstractPlant vacuolar H+-translocating inorganic pyrophosphatase (V-PPase EC 3.6. 1. 1) utilizes inorganic pyrophosphate (PP) as an energy source to generate a H+ gradient potential for the secondary transport of ions and metabolites across the vacuole membrane. In this study, functional roles of arginine residues in mung bean V-PPase were determined by site-directed mutagenesis. Alignment of amino-acid sequence of K+-dependent V-PPases from several organisms showed that I I of all 15 arginine residues were highly conserved. Arginine residues were individually substituted by alanine residues to produce R-A-substituted V-PPases, which were then heterologously expressed in yeast. The characteristics of mutant variants were subsequently scrutinized. As a result, most R -> A-substituted V-PPases exhibited similar enzymatic activities to the wild-type with exception that R242A, R523A, and R609A mutants markedly lost their abilities of PPi hydrolysis and associated W-translocation. Moreover, mutation on these three arginines altered the optimal pH and significantly reduced K+-stimulation for enzymatic activities, implying a conformational change or a modification in enzymatic reaction upon substitution. In particular, R242A performed striking resistance to specific arginine-modifiers, 2,3-butanedione and phenylglyoxal, revealing that Arg 242 is most likely the primary target residue for these two reagents. The mutation at Arg 242 also removed F- inhibition that is presumably derived from the interfering in the formation of substrate complex Mg2+-PPi. Our results suggest accordingly that active pocket of V-PPase probably contains the essential Arg 242 which is embedded in a more hydrophobic environment. (C) 2007 Elsevier B.V. All rights reserved.en_US
dc.language.isoen_USzh_TW
dc.relationBiochimica Et Biophysica Acta-Bioenergeticsen_US
dc.relation.ispartofseriesBiochimica Et Biophysica Acta-Bioenergetics, Volume 1767, Issue 7, Page(s) 965-973.en_US
dc.relation.urihttp://dx.doi.org/10.1016/j.bbabio.2007.04.007en_US
dc.subjectproton translocationen_US
dc.subjecttonoplasten_US
dc.subjectvacuoleen_US
dc.subjectsite-directed mutagenesisen_US
dc.subjectvacuolar H+-pyrophosphataseen_US
dc.subjecttranslocating inorganic pyrophosphataseen_US
dc.subjectinhibitionen_US
dc.subjectsiteen_US
dc.subjectarabidopsisen_US
dc.subjectmutationen_US
dc.subjectbindingen_US
dc.subjectmutagenesisen_US
dc.subjectexpressionen_US
dc.subjecttransporten_US
dc.subjectmembranesen_US
dc.titleFunctional roles of arginine residues in mung bean vacuolar H+-pyrophosphataseen_US
dc.typeJournal Articlezh_TW
dc.identifier.doi10.1016/j.bbabio.2007.04.007zh_TW
item.grantfulltextnone-
item.openairetypeJournal Article-
item.cerifentitytypePublications-
item.languageiso639-1en_US-
item.fulltextno fulltext-
item.openairecristypehttp://purl.org/coar/resource_type/c_18cf-
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