EFFECT OF GUANYL NUCLEOTIDES ON PARATHYROID HORMONE-RESPONSIVE ADENYLATE CYCLASE IN CHICK KIDNEY

Abstract

SUMMARY Both guanosine 5′-triphosphate (GTP) and 5′-guanylylimidodiphosphate (Gpp(NH)p) activated adenylate cyclase (EC 4.6.1.1) in chick kidney plasma membranes. Half-maximal stimulation occurred at 3·1 × 10⁻⁶ m for both agents. The maximum increases in adenylate cyclase activity produced by GTP and Gpp(NH)p were respectively 130 and 720% over basal activity. At the end of a 12 min incubation period GTP concentration was 85% of that originally added in the presence of an ATP-regenerating system but less than 20% in its absence. GTP and guanosine 5′-diphosphate inhibited the activation of adenylate cyclase by Gpp(NH)p, suggesting that they all acted at a common site. Gpp(NH)p facilitated the stimulation of adenylate cyclase activity by bovine parathyroid hormone (BPTH) and by the synthetic amino terminal fragment BPTH (1–34), decreasing the concentrations required for half-maximal enzyme activation by a factor of approximately eight in both cases. This property was not shared by the native nucleotide GTP. Gpp(NH)p rendered active (at certain concentrations) a synthetic parathyroid hormone peptide fragment, BPTH (2–34), which was incapable of activating adenylate cyclase in the absence of the nucleotide analogue. This suggested that the GTP analogue, in addition to a direct effect upon adenylate cyclase activity, was capable of influencing hormone interaction with the enzyme complex.