A model for the regulation of brain adenylate cyclase by ionic equilibria.

Multiple-equilibrium equations were solved to investigate the individual and separate effects of Mg2+, Mn2+, Ca2+, ATP4-, and their complexes on the kinetics of brain adenylate cyclase. The effects of divalent metals and/or ATP4- (in excess of their participation in complex formation) were determined and, from the corresponding apparent affinity values, the following kinetic constants were obtained: Km(MgATP) = 1.0 mM, Ki(ATP4-) = 0.27 mM, Km(MnATP) = 0.07 mM, and Ki(CaATP) = 0.015 mM. MgATP, MnATP, ATP4-, and CaATP were shown to compete for the active site of the enzyme. Hence, it is proposed that endogenous metabolites with a strong ligand activity for divalent metals, such as citrate and some amino acids, become integrated into a metabolite feedback control of the enzyme through the release of ATP4- from MgATP. Ca2+ fluxes may participate in the endogenous regulation of adenylate cyclase by modifying the level of CaATP. The free divalent metals show an order of affinity K0.5(Ca2+) = 0.02 mM, K0.5(Mn2+) = 3.8 mM, K0.5(Mg2+) - 4.7 mM, and an order of activity Mn2+ greater than Mg2+ greater than Ca2+. The data indicate that Mn2+ and Mg2+ ions may compete for a regulatory site distinct from the active site and increase Vm without changing Km(MgATP), Km(MnATP), or Ki(ATP4-). The interactions of ATP4- and CaATP, which act as competitive inhibitors of the reaction of the enzyme with the substrates MgATP and MnATP, and Mg2+ and Mn2+, which act as activators of the enzyme in the absence of hormones, are shown to follow the random rapid equilibrium BiBi group-transfer mechanism of Cleland with the stipulation that neither Mg2+ nor Mn2+, in excess of their respective participation in substrate formation, are obligatorily required for basal activity. ATP4- and CaATP are involved in dead-end inhibition. For MgCl2 saturation curves at constant total ATP concentration, the computer-generated curves based on the RARE BiBi model predict a change in the Hill cooperativity h from a basal value of 2.6, when Mg2+ is not obligatorily required, to 4.0 when the addition of hormones or neurotransmitters induces an obligatory requirement for Mg2+.

The hypothalamic-pituitary-gonadal axis during hyperthyroidism in the rat.

The rat, an animal without testosterone-estrogen-binding-globulin, was treated with L-T4 to the point of hyperthyroidism in order to study the hypothalamic-pituitary-testicular axis during this condition. Hyperthyroidism led to a significant decline in serum FSH, a fall in serum LH which was not satistically significant, and no change in serum levels of testosterone or estradiol. Testes of hyperthyroid rats produced significantly more testosterone during in vitro incubations than did the testes of control animals. We conclude that hyperthyroidism in the rat leads to a fall in FSH levels either via direct pituitary suppression or via accelerated FSH metabolism. In addition, in vitro studies suggest that excess thyroid hormone may stimulate intratesticular 17 beta-hydroxysteroid dehydrogenase.

Effects of noradrenaline on the activation and the stability of brain adenylate cyclase.

At constant 1 mM-ATP, the Mg2+-saturation curves for adenylate cyclase (EC 4.6.1.1) particulate preparations obtained from corpus striatum and cortex tissues of rat brain show that addition of 0.1 mM-noradrenaline increases the apparent Vmax. for Mg2+ by 300% in corpus striatum particles, and by 280% in cortex particles. At 10 mM-MgCl2, the addition of 0.1 mM-noradrenaline increased by 800% the adenylate cyclase activity of corpus striatum particles. At all Mg2+ concentrations, the addition of 0.3 mM-CaCl2 suppressed the noradrenaline-induced stimulation of adenylate cyclase of corpus striatum particles, and even resulted in a strong inhibition of the activating effect of Mg2+ itself on adenylate cyclase of corpus striatum particles, and even resulted in a strong inhibition of the activating effect of Mg2+ itself on adenylate cyclase activity of cortex particles. The addition of noradrenaline during a 3 h preincubation of particle preparations of brain cortex at 38 degrees C decreased by more than 4-fold the half-life of the decay of adenylate cyclase activity. The addition of MgATP protected against noradrenaline-induced inactivation.