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Conditionally expressed Gα_(15) couples to endogenous receptors in GH_3 cells

Offermanns, Stefan and Negulescu, Paul and Hu, Yi-Hui and Simon, Melvin I. (2001) Conditionally expressed Gα_(15) couples to endogenous receptors in GH_3 cells. Naunyn-Schmiedeberg's Archives of Pharmacology, 364 (2). pp. 140-148. ISSN 0028-1298. https://resolver.caltech.edu/CaltechAUTHORS:20191009-103456382

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Abstract

The mammalian G proteins G_(15) and G_(16) couple a wide variety of receptors to phospholipase C (PLC) in co-transfected systems, and it has been suggested that they can be used as tools in agonist-screening systems. Using the reversed tetracycline-controlled transactivation system we generated rat pituitary GH_3 cell clones that expressed Gα_(15) and Gα_(16) conditionally to study the coupling of endogenous receptors to both G proteins. In cells expressing moderate levels of Gα_(15), activation of various endogenous receptors increased inositol phosphate production, whereas conditional expression of Gα_(16) had no significant effect on agonist-dependent PLC activity. Activation of PLC through Gα_(15) in response to carbachol did not increase cytosolic [Ca2+] ([Ca^(2+)]_i) but stimulated protein kinase C. While carbachol decreased the secretory activity in non-induced GH_3 cells, it increased secretion in cells expressing Gα_(15). Our data demonstrate that Gα_(15) has a higher functional promiscuity than Gα_(16) when studied in a system that preserves physiological G protein and receptor levels. In addition, Gα_(15)-mediated coupling of a receptor to PLC can change the cellular response to receptor agonists, indicating that downstream cellular functions can be used to detect receptor activation in screening systems employing a promiscuous G protein.


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https://doi.org/10.1007/s002100100444DOIArticle
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Alternate Title:Conditionally expressed Gα15 couples to endogenous receptors in GH3 cells, Conditionally expressed G alpha 15 couples to endogenous receptors in GH3 cells
Additional Information:© 2001 Springer-Verlag. Received: 23 March 2001; Accepted: 19 April 2001; Published online: 8 June 2001. This work was supported by grants from the National Institutes of General Medical Sciences (GM 34236), from the National Institutes of Mental Health (MH 49176) and from the Deutsche Forschungsgemeinschaft.
Funders:
Funding AgencyGrant Number
NIHGM 34236
NIHMH 49176
Deutsche Forschungsgemeinschaft (DFG)UNSPECIFIED
Subject Keywords:G-protein; G15/G16; Promiscuous coupling; GH3 cells; Tet-system
Issue or Number:2
Record Number:CaltechAUTHORS:20191009-103456382
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20191009-103456382
Official Citation:Offermanns, S., Negulescu, P., Hu, YH. et al. Naunyn-Schmied Arch Pharmacol (2001) 364: 140. https://doi.org/10.1007/s002100100444
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:99187
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:09 Oct 2019 17:47
Last Modified:09 Oct 2019 17:47

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