Functional characterization of heterotrimeric G-proteins in rat diaphragm muscle

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dc.contributor.author Andrade-Lopes, Ana Luiza [UNIFESP]
dc.contributor.author Pires-Oliveira, Marcelo [UNIFESP]
dc.contributor.author Sandro Menezes-Rodrigues, Francisco [UNIFESP]
dc.contributor.author Godinho, Rosely Oliveira [UNIFESP]
dc.date.accessioned 2016-01-24T14:06:12Z
dc.date.available 2016-01-24T14:06:12Z
dc.date.issued 2011-02-15
dc.identifier http://dx.doi.org/10.1016/j.resp.2010.11.005
dc.identifier.citation Respiratory Physiology & Neurobiology. Amsterdam: Elsevier B.V., v. 175, n. 2, p. 212-219, 2011.
dc.identifier.issn 1569-9048
dc.identifier.uri http://repositorio.unifesp.br/handle/11600/33469
dc.description.abstract Seven-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.sponsorship Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.description.sponsorship Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
dc.format.extent 212-219
dc.language.iso eng
dc.publisher Elsevier B.V.
dc.relation.ispartof Respiratory Physiology & Neurobiology
dc.rights Acesso restrito
dc.subject Diaphragm en
dc.subject Skeletal muscle en
dc.subject G-protein en
dc.subject [(35)S]GTP gamma S en
dc.subject GPCR en
dc.subject Myoblast culture en
dc.subject L6 cell line en
dc.title Functional characterization of heterotrimeric G-proteins in rat diaphragm muscle en
dc.type Artigo
dc.rights.license http://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy
dc.contributor.institution Universidade Federal de São Paulo (UNIFESP)
dc.description.affiliation Universidade Federal de São Paulo UNIFESP, Dept Pharmacol, Div Cellular Pharmacol, BR-04044020 São Paulo, Brazil
dc.description.affiliationUnifesp Universidade Federal de São Paulo UNIFESP, Dept Pharmacol, Div Cellular Pharmacol, BR-04044020 São Paulo, Brazil
dc.description.sponsorshipID FAPESP: 05/59006-1
dc.identifier.doi 10.1016/j.resp.2010.11.005
dc.description.source Web of Science
dc.identifier.wos WOS:000287546200003



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