Identification and Quantitation of G-Protein α-Subunits

  • Ian Mullaney
  • Graeme Milligan
Part of the Methods in Molecular Biology book series (MIMB, volume 259)


The demonstration that many intracellular signaling processes are mediated by a family of closely related guanine nucleotide binding proteins (G-proteins) has led to the development of specific techniques that can be used to identify which of these polypeptide(s) is involved on receptor activation by ligand. In addition, these methods can be used to probe the specificity of the interaction and to yield information about the stoichiometries involved.

Key Words

G-protein immunoblotting pertussis toxin 


  1. 1.
    Goldsmith, P., Gierschik, P., Milligan, G., et al. (1987) Antibodies directed against synthetic peptides distinguish between GTP-binding proteins in neutophil and brain. J. Biol. Chem. 262, 14,683–14,688.PubMedGoogle Scholar
  2. 2.
    Milligan, G. (1993) Regional distribution and quantitative measurement of the phosphoinositidase C-linked guanine nucleotide binding proteins G11α and Gqα in rat brain. J. Neurochem. 61, 845–851.PubMedCrossRefGoogle Scholar
  3. 3.
    Wise, A., Lee, T. W., MacEwan, D. J., and Milligan, G. (1995) Degradation of G11α/Gqα is accelerated by agonist occupancy of α1A/D, α1B and α1C adrenergic receptors. J. Biol. Chem. 270, 17,196–17,203.PubMedCrossRefGoogle Scholar
  4. 4.
    Wise, A. and Milligan, G. (1994) High level expression of mammalian G-protein a subunit Gq subtypes in Escherichia coli. Biochem. Soc. Trans. 22, 12S.Google Scholar
  5. 5.
    Koski, G. and Klee, W. A. (1981) Opiates inhibit adenylate cyclase by stimulating GTP hydrolysis. Proc. Natl. Acad. Sci. USA 78, 4185–4189.PubMedCrossRefGoogle Scholar
  6. 6.
    Hudson, T. H. and Johnson, G. L. (1980) Peptide mapping of adenylyl cyclase regulatory proteins that are cholera toxin substrates. J. Biol. Chem. 255, 7480–7486.PubMedGoogle Scholar
  7. 7.
    Laemmli, U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–695.PubMedCrossRefGoogle Scholar
  8. 8.
    Towbin, H., Staehelin, T., and Gordon, J. (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc. Natl. Acad. Sci. USA 76, 4350–4354.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc. 2004

Authors and Affiliations

  • Ian Mullaney
    • 1
  • Graeme Milligan
    • 1
  1. 1.Molecular Pharmacology GroupUniversity of GlasgowGlasgowScotland, UK

Personalised recommendations