Immunological Probes of the Structure, Function, and Expression of Heterotrimeric GTP-Binding Proteins

  • Allen M. Spiegel
  • Paul K. Goldsmith
  • William F. Simonds
  • Teresa Jones
  • Kevin Rossiter
  • Cecilia G. Unson
Part of the Applications of Molecular Genetics to Pharmacology book series


G-proteins convey extracellular signals from cell surface receptors to diverse effectors such as enzymes of second messenger metabolism and ion channels. Receptor-effector coupling G-proteins are composed of three subunits, α, β, and γ, each separate gene products. Multiple forms of each type of subunit have been identified at the DNA and protein level (Gilman, 1987; Lochrie and Simon, 1988; Spiegel, 1987). Historically, α subunits were first identified by GTP affinity labeling and bacterial toxin-catalyzed ADP-ribosylation. These methods lack requisite specificity. Pertussis toxin, for example, is now known to covalently modify as many as six different Gα subunits (Gilman, 1987; Lochrie and Simon, 1988; Spiegel, 1987). Antibodies specific for G-protein subunits have proved to be powerful tools for studies of G-proteins. Both polyclonal (Gierschik et al., 1985, 1986c; Huff et al., 1985; Katada et al., 1987; Pines et al., 1985a; Roof et al., 1985) and monoclonal (Halpern et al., 1986; Hamm and Bownds, 1984; Navon and Fung, 1987, 1988; Tsai et al., 1987) antibodies have been raised against purified G-proteins. Antisera raised against purified G-holoproteins and subunits provided initial evidence for similarities (β subunits) and differences (α and γ subunits) between members of the G-protein family (Figure 11. 1). Immunochemical studies with such antisera suggested diversity in pertussis toxin substrates (Gierschik et al., 1986b), subsequently confirmed by cloning of cDNAs encoding multiple types of putative pertussis toxin substrate (Gilman, 1987; Lochrie and Simon, 1988; Spiegel, 1987).


Pertussis Toxin Immunochemical Study Peptide Antiserum Human Platelet Membrane Inhibitory Guanine Nucleotide 


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Copyright information

© Birkhäuser Boston 1992

Authors and Affiliations

  • Allen M. Spiegel
  • Paul K. Goldsmith
  • William F. Simonds
  • Teresa Jones
  • Kevin Rossiter
  • Cecilia G. Unson

There are no affiliations available

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