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  Gs couples thyrotropin-releasing hormone receptors expressed in Xenopus oocytes to phospholipase C

de la Peña, P., del Camino, D., Pardo, L. A., Dominguez, P., & Barros, F. (1995). Gs couples thyrotropin-releasing hormone receptors expressed in Xenopus oocytes to phospholipase C. Journal of Biological Chemistry, 270(8), 3554-9. doi:10.1074/jbc.270.8.3554.

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de la Peña, P.1, Author
del Camino, D.1, Author
Pardo, L. A.1, Author
Dominguez, P.1, Author
Barros, F.1, Author
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1Max Planck Society, ou_persistent13              

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Free keywords: Animals Base Sequence Calcium/metabolism Cell Line Chlorides/metabolism Enzyme Activation GTP-Binding Proteins/*metabolism Molecular Sequence Data Oligonucleotides, Antisense Oocytes RNA, Complementary Receptors, Serotonin/genetics Receptors, Thyrotropin-Releasing Hormone/genetics/*metabolism Recombinant Proteins/metabolism Type C Phospholipases/*metabolism Xenopus
 Abstract: Coupling of thyrotropin-releasing hormone (TRH) receptors to individual G-proteins has been studied in Xenopus oocytes injected with receptor cRNA and antisense oligonucleotides to mRNA encoding different G-protein alpha- and beta-subunits. Injection of antisenses which target mRNA sequences shared by several G-protein alpha or beta gamma polypeptides effectively blocked Ca(2+)-dependent Cl- currents induced by TRH through activation of phospholipase C. Three different alpha s-specific antisense oligonucleotides complementary to sequences located in different positions along the coding region of the alpha s protein mRNA were highly effective in inhibiting TRH-induced responses. Anti-alpha o, -alpha q, -alpha i, or -alpha z oligonucleotides were not able to modify the TRH-evoked response. In contrast, anti-alpha o, but not anti-alpha s, oligonucleotides blocked the response to serotonin in oocytes injected with serotonin 5-HT1c receptor cRNA. Cholera toxin catalyzed the [32P]ADP-ribosylation of 40-42- and 50-52-kDa proteins in GH3 cell plasma membranes. [32P]ADP-ribosylation of oocyte membranes with the toxin labeled several proteins. These include a single 50-55-kDa substrate, which is clearly diminished in membranes from anti-alpha s-injected oocytes. Amplification of oocyte RNA in a polymerase chain reaction system and sequencing of the amplified products demonstrated that anti-alpha s oligonucleotides selectively recognize the message for the Xenopus alpha s polypeptide. It is concluded that Gs, but not Go, Gq, Gi, or Gz, couples TRH receptors expressed in oocytes to activation of phospholipase C and subsequent inositol 1,4,5-trisphosphate-dependent stimulation of Ca(2+)-dependent Cl- currents.

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 Dates: 1995-02-241995
 Publication Status: Published in print
 Pages: -
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 Table of Contents: -
 Rev. Type: -
 Identifiers: Other: 7876090
DOI: 10.1074/jbc.270.8.3554
ISSN: 0021-9258 (Print) 0021-9258 (Linking)
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Title: Journal of Biological Chemistry
Source Genre: Journal
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Pages: - Volume / Issue: 270 (8) Sequence Number: - Start / End Page: 3554 - 9 Identifier: -