Nucleotide cyclases activation

This toxin subunit is an enzyme, an ADP-ribo-syltransferase which catalyzes transfer of ADP-ribosyl units from the coenzyme NAD+ to specific arginine side chains to form N-ADP-ribosyl derivatives of various proteins. Of the proteins modified by cholera toxin, the most significant is the guanyl nucleotide regulatory protein Gs of the adenylate cyclase system.C/f/h ADP ribosylation of arginine 201 of the a subunit of protein Gs inhibits the GTP hydrolysis that normally allows the protein to relax to an unactivated form.e The ADP-ribosylated Gs keeps adenylate cyclase activated continuously and... 

Wieczorek H. and Schweikl H. (1985) Concentrations of cyclic nucleotids and activities of cyclases and phosphodiesterases in an insect chemosensory organ. Insect Biochem. 6, 723-728. 

In other cell types, guanine nucleotides interact with a guanine nucleotide subunit (G- or Ng-subunit) to translate receptor stimulation into increased adenylate cyclase activity (12.) Cholera toxin inhibits a specific GTPase on this guanine nucleotide subunit and thereby increases adenylate cyclase activity (13.). In dispersed cells from the bovine parathyroid gland, cholera toxin markedly increases cAMP formation and causes a 3 to 10-fold increase in the apparent affinity cf dopamine for its receptor (as determined by cAMP accumulation or IR-PTH secretion (J y.). The effects of guanine nucleotides and cholera toxin on cAMP accumulation in parathyroid cells result from interactions with the guanine nucleotide subunit in this cell. 

The role of cyclic AMP as modulator of prolactin secretion was first suggested by the finding of a stimulatory effect of cyclic AMP derivatives (17-22) and inhibitors of cyclic nucleotide phosphodiesterase activity such as theophylline and IBMX (22-26) on the secretion of this hormone. More convincing evidence supporting a role of cyclic AMP in the action of dopamine on prolactin secretion had to be obtained, however, by measurement of adenohypophysial adenylate cyclase activity or cyclic AMP accumulation under the influence of the catecholamine. As illustrated in Fig. 1, addition of 100 nM dopamine to male rat hemipituitaries led to a rapid inhibition of cyclic AMP accumulation, a maximal effect (30% inhibition) being already obtained 5 min after addition of the catecholamine. Thus, while dopamine is well known to stimulate adenylate cyclase activity in the striatum (27, 28), its effect at the adenohypophysial level in intact cells is inhibitory. Dopamine has also been found to exert parallel inhibitory effects on cyclic AMP levels and prolactin release in ovine adenohypophysial cells in culture (29) and purified rat mammotrophs (30). Using paired hemipituitaries obtained from female rats, Ray and Wallis (22) have found a rapid inhibitory effect of dopamine on cyclic AMP accumulation to approximately 75% of control. 

The above-described data show that CRF added to cells of the rat Intermediate lobe In culture causes a rapid stimulation of oe-MSH release and cyclic AMP accumulation, thus demonstrating a direct action of the peptide on pars intermedia cells (15). It is however difficult, using intact cells, to dissociate between increases in cyclic AMP levels due to stimulation of adenylate cyclase activity or to Inhibition of cyclic nucleotide phosphodiesterase or to a combination of both effects. Definitive proof of the role of adenylate cyclase In the action of CRF In the intermediate lobe of the pituitary gland is provided by the following findings of a CRF-lnduced stimulation of adenylate cyclase activity in homogenate of rat and bovine pars Intermedia cells. 

As Illustrated in Fig. 7, 3 yM CRF and 1 yM (-)Isoproterenol cause a 190 and 110% stimulation of adenylate cyclase activity In rat pars intermedia particulate fraction, respectively. An additive effect Is observed when both stimulatory agents are present. Dopamine (30 yM), on the other hand, has no significant effect alone. However, In the presence of GXP, the catecholamine causes a 40 to 60% Inhibition of adenylate cyclase activity stimulated by CRF, ISO or CRF + ISO. It can also be seen that while 0.3 mM GXP alone causes a 100% increase In basal adenylate cyclase activity, it leads to a marked potentiation of the effect of ISO and CRF on [ 2P] cyclic AMP accumulation. It should be noticed that In the absence of the guanyl nucleotide, dopamine has no Inhibitory effect on adenylate cyclase activity In any of the groups studied. 

As mentioned earlier, guanyl nucleotides have been found to play an important role In the activation of adenylate cyclase activity by many hormones (90, 91). The present observations show that in pars Intermedia tissue, GXP causes an almost doubling of the stimulatory effect of CRF while that of the B-adrenerglc ago-... 

However, the levels of Gsa, Gia-1, Gia-2, Gia-3, Goa, and Gp were also shown to be unaltered in myocardium from SHRs, and adenylyl cyclase activity stimulated by PGEi, glucagon, and isoproterenol was reduced in SHRs, whereas FSK-stimulated enzyme activity was greater in SHRs as compared to WKY (McLellan et al. 1993). On the other hand, a diminished stimulation of adenylyl cyclase by stimulatory hormones, guanine nucleotides, FSK, and NaF in aorta and heart sarcolemma from SHRs (Anand-Srivastava 1992), renal hypertensive rats (Anand-Srivastava 1988) 1K1C HRs (Ge et al. 1999, 2006), and DOCA-salt HRs (Anand-Srivastava et al. 1993) has been demonstrated The reduction in the hormone receptor binding sites may be one of the possible mechanisms responsible for such an impaired response of hormones (Limas and Limas 1978 Woodcock et al. 

Anand-Srivastava MB. 1993. Rat platelets from spontaneously hypertensive rats exhibit decreased expression of inhibitory guanine nucleotide regulatory protein relationship with adenylate cyclase activity. Circ Res 173 1032-1039. 

Ikegaya, T., Kobayashi, A., Hong, R.B., Masuda, H., Kaneko, M., and Noboru, Y. 1992. Stimulatory guanine nucleotide-binding protein and adenylate cyclase activities in Bio 14.6 cardiomyopathic hamsters at the hypertrophic stage. Mol. Cell. Biochem. 110 83-90. 

Earlier experiments have shown that cholate extracts from transformed lung fibroblasts having only the Mr 52000 subunit of Gs are active in reconstituting hormone-, nucleotide- and fluoride-stimulated adenylyl cyclase activities in cyc membranes [178]. Human erythrocyte Gs, which has only the A/r 42000 a subunit(s) [22,179], also reconstitute(s) these functions. After partial separation, rabbit liver p52 Gs appeared to reconstitute hormone-stimulated activity better than p45 Gs... 

A variety of guanine nucleotide binding proteins (G-proteins) involved with the regulation of adenylate cyclase activity and transducin in the retina (Section 2.3.1) are substrates for ADP-ribosylation. Cholera toxin and E. coli enterotoxin LT ADP-ribosylate, and hence activate, the stimulatory G-protein of adenylate cyclase, whereas pertussis toxin ADP-ribosylates, and inactivates the inhibitory G-protein of adenylate cyclase. The result of ADP-ribosylation by either mechanism is increased adenylate cyclase activity, and an increase in intracellular cAMP and the opening of membrane calcium channels. Again, there are endogenous ADP-ribosyltransferases that modify the same G-proteins, but in a controlled manner (Moss et al., 1997, 1999). 

Nucleotide Cyclases. Calmodulin regulates the activity of different nucleotide cyclases such as adenylyl cyclase that catalyzes the formation of cAMP from ATP. cAMP is another important molecule in the cell functioning as a second-messenger like calcium. The cahnodulin-adenylyl cyclase interaction links these two second messengers together. ... 

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