Publication
Decavanadate binding to a high affinity site near the myosin catalytic centre inhibits F-actin-stimulated myosin ATPase activity
| dc.contributor.author | Tiago, Teresa | |
| dc.contributor.author | Aureliano, M. | |
| dc.contributor.author | Gutiérrez-Merino, Carlos | |
| dc.date.accessioned | 2012-06-28T08:10:24Z | |
| dc.date.available | 2012-06-28T08:10:24Z | |
| dc.date.issued | 2004 | |
| dc.description.abstract | Decameric vanadate (V10) inhibits the actin-stimulated myosin ATPase activity, noncompetitively with actin or with ATP upon interaction with a high-affinity binding site (Ki ) 0.27 ( 0.05 íM) in myosin subfragment-1 (S1). The binding of V10 to S1 can be monitored from titration with V10 of the fluorescence of S1 labeled at Cys-707 and Cys-697 with N-iodo-acetyl-N¢-(5-sulfo-1-naphthyl)- ethylenediamine (IAEDANS) or 5-(iodoacetamido) fluorescein, which showed the presence of only one V10 binding site per monomer with a dissociation constant of 0.16-0.7 íM, indicating that S1 labeling with these dyes produced only a small distortion of the V10 binding site. The large quenching of AEDANSlabeled S1 fluorescence produced by V10 indicated that the V10 binding site is close to Cys-697 and 707. Fluorescence studies demonstrated the following: (i) the binding of V10 to S1 is not competitive either with actin or with ADPâV1 or ADPâAlF4; (ii) the affinity of V10 for the complex S1/ADPâV1 and S1/ ADPâAlF4 is 2- and 3-fold lower than for S1; and (iii) it is competitive with the S1 “back door” ligand P1P5-diadenosine pentaphosphate. A local conformational change in S1 upon binding of V10 is supported by (i) a decrease of the efficiency of fluorescence energy transfer between eosin-labeled F-actin and fluorescein-labeled S1, and (ii) slower reassociation between S1 and F-actin after ATP hydrolysis. The results are consistent with binding of V10 to the Walker A motif of ABC ATPases, which in S1 corresponds to conserved regions of the P-loop which form part of the phosphate tube. | por |
| dc.identifier.issn | 0006-2960 | |
| dc.identifier.uri | http://hdl.handle.net/10400.1/1358 | |
| dc.language.iso | eng | por |
| dc.peerreviewed | yes | por |
| dc.publisher | American Chemical Society | por |
| dc.subject | Myosin | por |
| dc.subject | Decavanadate | por |
| dc.title | Decavanadate binding to a high affinity site near the myosin catalytic centre inhibits F-actin-stimulated myosin ATPase activity | por |
| dc.type | journal article | |
| dspace.entity.type | Publication | |
| oaire.citation.endPage | 5561 | por |
| oaire.citation.issue | 43 | por |
| oaire.citation.startPage | 5551 | por |
| oaire.citation.title | Biochemistry | por |
| person.familyName | Aureliano | |
| person.familyName | Gutierrez-Merino | |
| person.givenName | Manuel | |
| person.givenName | Carlos | |
| person.identifier | 584146 | |
| person.identifier.ciencia-id | AA14-3490-DC5E | |
| person.identifier.orcid | 0000-0003-4858-3201 | |
| person.identifier.orcid | 0000-0003-3673-7007 | |
| person.identifier.rid | I-3283-2012 | |
| person.identifier.rid | K-4574-2014 | |
| person.identifier.scopus-author-id | 6603412860 | |
| rcaap.rights | openAccess | por |
| rcaap.type | article | por |
| relation.isAuthorOfPublication | bb413661-7edd-4b57-8338-33889cfd05db | |
| relation.isAuthorOfPublication | 1edd3bda-28a9-4d7f-a404-f867f72ddefa | |
| relation.isAuthorOfPublication.latestForDiscovery | 1edd3bda-28a9-4d7f-a404-f867f72ddefa |