The possible interaction of bombesin receptors with guanine nucleotide binding protein in guinea pig lung was studied. The non-hydrolysable GTP analogue guanosine-5'-[gamma-thio]triphosphate (GTP gamma S) was shown to decrease [125I-Tyr4]bombesin binding in a concentration-dependent manner. The specificity of this effect was assessed by examining the effects of other guanine nucleotides on this binding at a concentration of 1 mM. GMP and GDP weakly inhibited [125I-Tyr4]bombesin binding (2 and 19%, respectively), whereas GTP, guanosine-5'-[beta-thio]triphosphate (GDP beta S), and 5-guanylylimidodiphosphate (GppNHp) exhibited similar potencies, inducing 52%, 46%, and 43% inhibition of [125I-Tyr4]bombesin binding respectively. Saturation experiments performed in the absence and presence of 100 microM GTP gamma S indicated the presence of a single population of receptors in both cases. However, the addition of GTP gamma S induced a marked decrease in the number of receptors (from 1.76 fmol/mg protein to 0.78 fmol/mg protein) without significantly altering the dissociation constant (Kd). These results provide evidence that bombesin receptors are coupled to a G-protein signal transduction pathway in guinea pig lung. We have further characterised this G-protein on the basis of its toxin sensitivity. Pretreatment of the lung membranes with either pertussis (10 micrograms/ml) or cholera toxin (50 micrograms/ml) was performed. Cholera toxin treatment did not affect the ability of GTP gamma S to inhibit [125I-Tyr4]bombesin binding to guinea pig lung membranes. However, pertussis toxin treatment induced a decrease in binding and resulted in the inability of GTP gamma S to inhibit [125I-Tyr4]bombesin binding in a concentration-dependent manner.(ABSTRACT TRUNCATED AT 250 WORDS)