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Receptors that couple to 2 classes of G proteins increase cAMP and activate CFTR expressed in Xenopus oocytes

Uezono, Y. and Bradley, J. and Min, C. and McCarty, N. A. and Quick, M. and Riordan, J. R. and Chavkin, C. and Zinn, K. and Lester, H. A. and Davidson, N. (1993) Receptors that couple to 2 classes of G proteins increase cAMP and activate CFTR expressed in Xenopus oocytes. Receptors & Channels, 1 (3). pp. 233-241. ISSN 1060-6823.

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The cystic fibrosis transmembrane conductance regulator (CFTR), a Cl- channel activated by phosphorylation, was expressed in Xenopus oocytes along with various combinations of several other components of the cAMP signalling pathway. Activation of the coexpressed beta 2 adrenergic receptor increased cAMP and led to CFTR activation. The activation of CFTR (1) requires only short (15 s) exposure to isoproterenol, (2) occurs for agonist concentrations 100-1000 fold lower than those that produce cAMP increases detectable by a radioimmunoassay, (3) requires injection of only 5 pg of receptor cRNA per oocyte, and (4) can be increased further by coexpression of cRNA for adenylyl cyclase type II or III or for Gs alpha. In addition, CFTR activation and cAMP increases by beta 2 activation were enhanced by activation of the coexpressed 5HT1A receptor, which is thought to couple to Gi. The additional activation by the 5HT1A receptor was enhanced by coexpression of adenylyl cyclase type II but not with type III and may proceed via the beta gamma subunits of a G protein. The sensitivity of the assay system is also demonstrated by responses to vasoactive intestinal peptide and to pituitary adenylate cyclase-activating polypeptide in oocytes injected with cerebral cortex mRNA.

Item Type:Article
Zinn, K.0000-0002-6706-5605
Lester, H. A.0000-0002-5470-5255
Additional Information:© 1993 Harwood Academic. (Received March 15, 1993; Revised May 15, 1993) We thank Dr.s M. Simon and Anna Aragay for advice and the gift of Gs cDNA, R. Reed for the gift of the adenyl cyclase type II cDNA, and Dr. B. Kobilka for the gift of the β2 receptor cDNA. We thank Drs. N. Dascal and C. Strader for comments on an earlier version of the manuscript. This work was supported by grants from the National Institutes of Health, the National Institute on Drug Abuse, the Silvio Conte Center for Neuroscience Research of the National Institute of Mental Health, the Cystic Fibrosis Foundation, the California Tobacco-Related Disease Program, the Wiersma Memorial Fund, and the Japanese Foundation for Clinical Pharmacology.
Funding AgencyGrant Number
National Institute on Drug Abuse (NIDA)UNSPECIFIED
National Institute of Mental Health (NIMH)UNSPECIFIED
Cystic Fibrosis FoundationUNSPECIFIED
California Tobacco-Related Disease Research ProgramUNSPECIFIED
Wiersma Memorial FundUNSPECIFIED
Japanese Foundation for Clinical PharmacologyUNSPECIFIED
Subject Keywords:Gs; Gi; cystic fibrosis; β2 receptor; serotonin receptors
Issue or Number:3
Record Number:CaltechAUTHORS:20151217-104726852
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:63038
Deposited By: George Porter
Deposited On:29 Jan 2016 00:11
Last Modified:03 Oct 2019 09:24

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