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Quenching of myosin intrinsic fluorescence unravels the existence of a high affinity binding site for decavanadate

dc.contributor.authorAureliano, M.
dc.date.accessioned2014-07-23T10:53:46Z
dc.date.available2014-07-23T10:53:46Z
dc.date.issued2002-03
dc.description.abstractDecavanadate, one of the aggregated species of vanadate, is a potent inhibitor of several enzymes, includingskeletalmusclemyosin.However,itsputativebindingsitesinmyosinarelargelyunknown. Titration of the intrinsic fluorescence of myosin, purified from rabbit skeletal muscle, have been carried out in 0.3 M KCl, 5 mM CaCl2 and 25 mM Tris-HCl (pH 7.0), with 0.1 mg/ml myosin. In the 0–200M total vanadate concentration range, decavanadate produced approximately 25% quenching of the intrinsic fluorescence of myosin, with an apparent dissociation constant in the micromolar range. This effect was found to be specific of decavanadate, because titration with metavanadate up to 200M did not produce a significant quenching of the intrinsic fluorescence of myosin. This quenching was accompanied by a parallel decrease of the accessibility of myosin tryptophans to the water-soluble collisional quencher KI, with an apparent dissociation constant also in the micromolar range. It is concluded that the binding of decavanadate to high-affinity sites in myosin produces local conformational change(s) near the tryptophans more accessible to water in the three-dimensional structure of this protein.por
dc.identifier.issn1053-0509
dc.identifier.otherAUT: MNU00233
dc.identifier.urihttp://hdl.handle.net/10400.1/4809
dc.language.isoengpor
dc.peerreviewedyespor
dc.publisherSpringer Verlagpor
dc.subjectMyosinpor
dc.subjectDecavanadatepor
dc.subjectSkeletal muscle myosinpor
dc.subjectIntrinsic fluorescencepor
dc.titleQuenching of myosin intrinsic fluorescence unravels the existence of a high affinity binding site for decavanadatepor
dc.typejournal article
dspace.entity.typePublication
oaire.citation.issue1por
oaire.citation.titleJournal of Fluorescencepor
oaire.citation.volume12por
person.familyNameAureliano
person.givenNameManuel
person.identifier584146
person.identifier.ciencia-idAA14-3490-DC5E
person.identifier.orcid0000-0003-4858-3201
person.identifier.ridI-3283-2012
person.identifier.scopus-author-id6603412860
rcaap.rightsrestrictedAccesspor
rcaap.typearticlepor
relation.isAuthorOfPublicationbb413661-7edd-4b57-8338-33889cfd05db
relation.isAuthorOfPublication.latestForDiscoverybb413661-7edd-4b57-8338-33889cfd05db

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