Linked Life Data

8175713

Source:http://linkedlifedata.com/resource/pubmed/id/8175713

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pubmed-article:8175713pubmed:abstractTextInsect cell expression systems are used to characterize signaling components such as G protein-coupled receptors. As such, one must know whether endogenous G proteins couple to non-native receptors. We examined G protein linkages after infection of Sporodoptera frugiperda (Sf9) cells with a baculovirus encoding the 5-HT1A receptor. Receptor expression was confirmed by immunoblot. Some of the receptors were functional, showing guanine nucleotide-sensitive binding to the specific agonist ligand [3H]8-hydroxy-2-(di-n-propylamino)-1,2,3,4-tetranaphthalene). Peak expression (approximately 150 fmol/mg of membrane protein) was attained approximately 72-96 h post-infection. 5-HT-increased covalent binding of [32P]GTP-azidoanilide to a 40 kDa band, which was identified as a G protein by nucleotide blocking, Mg2+ dependence, and immunoblot and immunoprecipitation studies. The band comigrated with 1) pertussis toxin substrate(s), and 2) a band recognized by two G(o) alpha antisera and one common to heterotrimeric G protein alpha-subunits, but not by sera specific for Gs alpha or G(i) alpha. Labeled species could be precipitated with a G(o) alpha antiserum. 5-HT-increased labeling of the band was prevented by preincubation with pertussis toxin. These studies suggest that the 5-HT1A receptor couples effectively to native insect cell G(o)-like proteins.lld:pubmed
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pubmed-article:8175713pubmed:authorpubmed-author:MulheronJ GJGlld:pubmed
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pubmed-article:8175713pubmed:authorpubmed-author:RaymondJ RJRlld:pubmed
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pubmed-article:8175713pubmed:pagination12954-62lld:pubmed
pubmed-article:8175713pubmed:dateRevised2007-11-14lld:pubmed
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pubmed-article:8175713pubmed:year1994lld:pubmed
pubmed-article:8175713pubmed:articleTitleHuman 5-HT1A receptor expressed in insect cells activates endogenous G(o)-like G protein(s).lld:pubmed
pubmed-article:8175713pubmed:affiliationDepartment of Medicine, Nephrology and Gastroenterology, Duke University Medical Center, Durham, North Carolina 27710.lld:pubmed
pubmed-article:8175713pubmed:publicationTypeJournal Articlelld:pubmed
pubmed-article:8175713pubmed:publicationTypeResearch Support, U.S. Gov't, P.H.S.lld:pubmed
pubmed-article:8175713pubmed:publicationTypeResearch Support, U.S. Gov't, Non-P.H.S.lld:pubmed
pubmed-article:8175713pubmed:publicationTypeResearch Support, Non-U.S. Gov'tlld:pubmed
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