Immunological identification of the α subunit of G13, a novel guanine nucleotide binding protein

Abstract

An antiserum (13CB) was generated against a synthetic peptide, HDNLKQLMLQ, which is predicted to represent the C-terminal decapeptide of the α subunit of the novel G-protein, G13. Competitive ELISA indicated that the antiserum reacted with this peptide but that it showed minimal ability to recognize peptides which represent the equivalent regions of the pertussis toxin-insensitive G-proteins, Gq + G11, G12, G15 + G16, G_L1 (also called G14) as Gz, and well as other G-proteins. Immunoblots of human platelet membranes with antiserum 13CB identified a single 43-kDa polypeptide, and while this immunoreactivity was abolished by the presence of the cognate peptide it was not modified by the presence of peptides corresponding to the equivalent region of other G-proteins. Immunoreactivity corresponding to G13α was detected in a range of cell types with human platelets having the highest levels of this polypeptide.