PMID: 2111013May 1, 1990Paper

2-Azido-[32P]NAD+, a photoactivatable probe for G-protein structure: evidence for holotransducin oligomers in which the ADP-ribosylated carboxyl terminus of alpha interacts with both alpha and gamma subunits

Proceedings of the National Academy of Sciences of the United States of America
R R VaillancourtA E Ruoho

Abstract

A radioactive and photoactivatable derivative of NAD+, 2-azido-[adenylate-32P]NAD+, has been synthesized and used with pertussis toxin to ADP-ribosylate Cys347 of the alpha subunit (alpha T) of GT, the retinal guanine nucleotide-binding protein. ADP-ribosylation of alpha T followed by light activation of the azide moiety of 2-azido-[adenylate-32P]ADP-ribose produced four crosslinked species involving the alpha and gamma subunits of the GT heterotrimer: an alpha trimer (alpha-alpha-alpha), and alpha-alpha-gamma crosslink, an alpha dimer (alpha-alpha), and an alpha-gamma crosslink. The alpha trimer, alpha-alpha-gamma complex, alpha dimer, and alpha-gamma complexes were immunoreactive with alpha T antibodies. The alpha-alpha-gamma and the alpha-gamma complexes were immunoreactive with antisera recognizing gamma subunits. No evidence was found for crosslinking of alpha T to beta T subunits. Hydrolysis of the thioglycosidic bond between Cys347 and 2-azido-[adenylate-32P]ADP-ribose using mercuric acetate resulted in the transfer of radiolabel from Cys347 of alpha T in the crosslinked oligomers to alpha monomers, indicative of intermolecular photocrosslinking, and to gamma monomers, indicative of either intermolecular crosslinked comp...Continue Reading

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Citations

Oct 1, 1991·Journal of Cellular Biochemistry·G L JohnsonA E Ruoho
Dec 1, 1991·Cellular and Molecular Neurobiology·H E Hamm
Dec 15, 2004·Biochemistry·Paul S-H ParkKrzysztof Palczewski
Aug 15, 1991·Proceedings of the National Academy of Sciences of the United States of America·S Nakamura, M Rodbell
Nov 1, 1993·Proceedings of the National Academy of Sciences of the United States of America·T C ThomasE J Neer
Jan 1, 1997·Annual Review of Biochemistry·S R Sprang
Feb 17, 1999·Proceedings of the National Academy of Sciences of the United States of America·T ExnerB Nürnberg
Dec 3, 2014·Current Opinion in Chemical Biology·Nathan P Westcott, Howard C Hang
Jun 3, 2004·The Journal of Biological Chemistry·Zhixian ZhangTheodore G Wensel
Nov 2, 2001·The Journal of Biological Chemistry·Valerie A MosserMichael I Schimerlik

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