Post-translational thio-acylation of a fusion protein between the α2A-adrenoceptor and the α subunit of the G protein Go1 is both dynamic and regulated by agonist binding. Incorporation of [3H]palmitate into the fusion protein was reduced substantially in the presence of the agonist adrenaline. This was dependent on the concentration of adrenaline and correlated with occupancy of the ligand binding site. Both the receptor and G-protein elements of the fusion construct incorporated [3H]palmitate but this occurred more rapidly for the G-protein element and regulation of acylation by the agonist occurred only for the G protein. The kinetics of de-palmitoylation of the α2A-adrenoceptor–Gαo1 fusion were accelerated markedly by agonist. Again, this reflected modulation of the G protein but not of the receptor. Agonist-induced regulation of the kinetics of thio-acylation of the G protein was abolished, however, in a mutant unable to bind guanosine 5′-[γ-[35S]thio]triphosphate ([35S]GTP[S]) in response to adrenaline. Despite the dynamic nature of the post-translational acylation and its regulation by agonist, the ability of adrenaline to activate the G protein, monitored by stimulation of the binding of [35S]GTP[S] to such fusion constructs, was unaffected by the palmitoylation potential of either the receptor or G-protein element.
Skip Nav Destination
Article navigation
January 2005
-
Cover Image
Cover Image
- PDF Icon PDF LinkFront Matter
- PDF Icon PDF LinkTable of Contents
- PDF Icon PDF LinkEditorial Board
Research Article|
December 14 2004
Activation of an α2A-adrenoceptor–Gαo1 fusion protein dynamically regulates the palmitoylation status of the G protein but not of the receptor
Elaine BARCLAY;
Elaine BARCLAY
*Molecular Pharmacology Group, Division of Biochemistry and Molecular Biology, University of Glasgow, Glasgow G12 8QQ, Scotland, U.K.
Search for other works by this author on:
Mark O'REILLY;
Mark O'REILLY
†Pfizer Global Research and Development, Sandwich, Kent, CT13 N9J, U.K.
Search for other works by this author on:
Graeme MILLIGAN
Graeme MILLIGAN
1
*Molecular Pharmacology Group, Division of Biochemistry and Molecular Biology, University of Glasgow, Glasgow G12 8QQ, Scotland, U.K.
1To whom correspondence should be addressed (email g.milligan@bio.gla.ac.uk).
Search for other works by this author on:
Publisher: Portland Press Ltd
Received:
August 20 2004
Revision Received:
September 01 2004
Accepted:
September 14 2004
Accepted Manuscript online:
September 14 2004
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London
2005
Biochem J (2005) 385 (1): 197–206.
Article history
Received:
August 20 2004
Revision Received:
September 01 2004
Accepted:
September 14 2004
Accepted Manuscript online:
September 14 2004
Citation
Elaine BARCLAY, Mark O'REILLY, Graeme MILLIGAN; Activation of an α2A-adrenoceptor–Gαo1 fusion protein dynamically regulates the palmitoylation status of the G protein but not of the receptor. Biochem J 1 January 2005; 385 (1): 197–206. doi: https://doi.org/10.1042/BJ20041432
Download citation file:
Sign in
Don't already have an account? Register
Sign in to your personal account
You could not be signed in. Please check your email address / username and password and try again.
Captcha Validation Error. Please try again.