Functional characterization of heterotrimeric G-proteins in rat diaphragm muscle

Página do item simplificado
dc.contributor.authorAndrade-Lopes, Ana Luiza [UNIFESP]
dc.contributor.authorPires-Oliveira, Marcelo [UNIFESP]
dc.contributor.authorSandro Menezes-Rodrigues, Francisco [UNIFESP]
dc.contributor.authorGodinho, Rosely Oliveira [UNIFESP]
dc.contributor.institutionUniversidade Federal de São Paulo (UNIFESP)
dc.date.accessioned2016-01-24T14:06:12Z
dc.date.available2016-01-24T14:06:12Z
dc.date.issued2011-02-15
dc.description.abstractSeven-transmembrane receptors mediate diverse skeletal muscle responses for a wide variety of stimuli, via activation of heterotrimeric G-proteins. Herein we evaluate the expression and activation of rat diaphragm or cultured skeletal muscle G-proteins using [(35)S]GTP gamma S. Total membrane G alpha subunit content was 4-7 times higher in rat primary cultured myotubes and L6 cell line than in diaphragm (32.6 +/- 1.2 fmol/mg protein) and 7-27% of them were in the active conformational state. Immunoprecipitation assay showed equal expression of diaphragm G alpha s, G alpha q and G alpha i/o. Addition of GDP allowed the measurement of G-protein activation by different GPCR, including adrenoceptor, adenosine, melatonin and muscarinic receptors. Diaphragm denervation resulted in a marked increase in both total and active state G-protein levels. Together, the results show that [(35)S]GTP gamma S binding assay is a sensitive and valuable method to evaluate GPCR activity in skeletal muscle cells, which is of particular interest for pharmacological analysis of drugs with potential use in the management of respiratory muscle failure. (C) 2010 Elsevier B.V. All rights reserved.en
dc.description.affiliationUniversidade Federal de São Paulo UNIFESP, Dept Pharmacol, Div Cellular Pharmacol, BR-04044020 São Paulo, Brazil
dc.description.affiliationUnifespUniversidade Federal de São Paulo UNIFESP, Dept Pharmacol, Div Cellular Pharmacol, BR-04044020 São Paulo, Brazil
dc.description.sourceWeb of Science
dc.description.sponsorshipFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.description.sponsorshipConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
dc.description.sponsorshipIDFAPESP: 05/59006-1
dc.format.extent212-219
dc.identifierhttp://dx.doi.org/10.1016/j.resp.2010.11.005
dc.identifier.citationRespiratory Physiology & Neurobiology. Amsterdam: Elsevier B.V., v. 175, n. 2, p. 212-219, 2011.
dc.identifier.doi10.1016/j.resp.2010.11.005
dc.identifier.issn1569-9048
dc.identifier.urihttp://repositorio.unifesp.br/handle/11600/33469
dc.identifier.wosWOS:000287546200003
dc.language.isoeng
dc.publisherElsevier B.V.
dc.relation.ispartofRespiratory Physiology & Neurobiology
dc.rightsinfo:eu-repo/semantics/restrictedAccess
dc.rights.licensehttp://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy
dc.subjectDiaphragmen
dc.subjectSkeletal muscleen
dc.subjectG-proteinen
dc.subject[(35)S]GTP gamma Sen
dc.subjectGPCRen
dc.subjectMyoblast cultureen
dc.subjectL6 cell lineen
dc.titleFunctional characterization of heterotrimeric G-proteins in rat diaphragm muscleen
dc.typeinfo:eu-repo/semantics/article
Arquivos
Coleções
EPM - Artigos