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Molecular Pharmacology

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Research ArticleArticle

Incorporation of Gαz-Specific Sequence at the Carboxyl Terminus Increases the Promiscuity of Gα16toward Gi-Coupled Receptors

Sejal M. Mody, Maurice K. C. Ho, Sushma A. Joshi and Yung H. Wong
Molecular Pharmacology January 2000, 57 (1) 13-23;
Sejal M. Mody
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Maurice K. C. Ho
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Sushma A. Joshi
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Yung H. Wong
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Abstract

Although the promiscuous nature of G16 allows it to interact with numerous G protein-coupled receptors, several Gi-linked receptors are incapable of activating phospholipase C via G16. A series of chimeras between Gα16 and Gαz were constructed and assayed for their ability to mediate receptor-induced stimulation of phospholipase C. Two Gα16/z chimeras harboring 25 or 44 Gαz-specific sequences at their C termini (named 16z25 and 16z44) were capable of responding to 14 different Gi-coupled receptors tested, including those that were either unable to associate with Gα16 (melatonin Mel1c) or activate Gα16 weakly (μ-opioid and type 1 somatostatin). Agonist-induced stimulation of phospholipase C was more efficiently mediated (higher maximal and lower EC50 value) by 16z44 than by Gα16. Both 16z25 and 16z44 were also coupled to Gs- and Gq-linked receptors. Incorporation of Gαz sequence at the N terminus of Gα16 did not further enhance the ability of the chimeras to interact with Gi-coupled receptors. Expression of the various chimeras was verified by immunodetection and functional analysis of their constitutively activated mutants. These results show that the incorporation of α4/β6 and α5 regions of Gαz into a Gα16 backbone can improve the recognition of Gi-coupled receptors. Gα16/zchimeras with expanded capability to interact with Gi-linked receptors may be used to link orphan receptors to the stimulation of phospholipase C.

Footnotes

  • Send reprint requests to: Dr. Yung H. Wong, Department of Biology, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong, China. E-mailboyung{at}ust.hk

  • ↵1 Present address: Center for Biotechnology, NorthWestern University, 1801 Maple Avenue, Evanston, IL 60201.

  • This work was supported by the Research Grants Council of Hong Kong (HKUST 6096/98M).

  • Abbreviations:
    GPCR
    G protein-coupled receptor
    DPDPE
    [d-Pen2,d-Pen5]enkephalin
    PLC
    phospholipase C
    PCR
    polymerase chain reaction
    PTX
    pertussis toxin
    IP
    inositol phosphates
    DMEM
    Dulbecco's modified Eagle's medium
    • Received April 22, 1999.
    • Accepted September 3, 1999.
  • The American Society for Pharmacology and Experimental Therapeutics
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Molecular Pharmacology: 57 (1)
Molecular Pharmacology
Vol. 57, Issue 1
1 Jan 2000
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Research ArticleArticle

Incorporation of Gαz-Specific Sequence at the Carboxyl Terminus Increases the Promiscuity of Gα16toward Gi-Coupled Receptors

Sejal M. Mody, Maurice K. C. Ho, Sushma A. Joshi and Yung H. Wong
Molecular Pharmacology January 1, 2000, 57 (1) 13-23;

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Research ArticleArticle

Incorporation of Gαz-Specific Sequence at the Carboxyl Terminus Increases the Promiscuity of Gα16toward Gi-Coupled Receptors

Sejal M. Mody, Maurice K. C. Ho, Sushma A. Joshi and Yung H. Wong
Molecular Pharmacology January 1, 2000, 57 (1) 13-23;
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