scholarly journals The local anaesthetic benzyl alcohol attenuates the α2-adrenoceptor-mediated inhibition of human platelet adenylate cyclase activity when stimulated by prostaglandin E1, but not that stimulated by forskolin

1989 ◽  
Vol 264 (2) ◽  
pp. 483-488 ◽  
Author(s):  
S Spence ◽  
M D Houslay
Keyword(s):  
Adenylate Cyclase ◽  
Benzyl Alcohol ◽  
Inhibitory Action ◽  
Prostaglandin E1 ◽  
Regulatory Protein ◽  
Inhibitory Response ◽  
Dose Effect ◽  
Cyclase Activity ◽  
Adenylate Cyclase Activity ◽  
Catalytic Unit

Treatment of human platelets with concentrations of benzyl alcohol up to 50 mM augmented adenylate cyclase activity when it was assayed in the basal state and when stimulated by prostaglandin E1 (PGE1), isoprenaline or NaF. Benzyl alcohol antagonized the stimulatory effect exerted on the catalytic unit of adenylate cyclase by the diterpene forskolin. Benzyl alcohol did not modify the magnitude of the inhibitory response when the catalytic unit of adenylate cyclase was inhibited by using either low concentrations of guanosine 5′-[beta gamma-imido]triphosphate, which acts selectively on the inhibitory guanine nucleotide-regulatory protein Gi, or during alpha 2-adrenoceptor occupancy, by using adrenaline (+ propranolol). Some 34% of the potent inhibitory action of adrenaline on PGE1-stimulated adenylate cyclase was obliterated in a dose-dependent fashion (concn. giving 50% inhibition = 12.5 mM) by benzyl alcohol, with the residual inhibitory action being apparently resistant to the action of benzyl alcohol at concentrations up to 50 mM. Treatment of membranes with benzyl alcohol did not lead to the release of either the alpha-subunit of Gi or G-protein subunits. The alpha 2-adrenoceptor-mediated inhibition of adenylate cyclase was abolished when assays were performed in the presence of Mn2+ rather than Mg2+ and, under such conditions, dose-effect curves for the action of benzyl alcohol on PGE1-stimulated adenylate cyclase activity were similar whether or not adrenaline (+propranolol) was present. We suggest that (i) alpha 2-adrenoceptor- and Gi-mediated inhibition of PGE1-stimulated adenylate cyclase may have two components, one of which is sensitive to inhibition by benzyl alcohol, and (ii) the Gi-mediated inhibition of forskolin-stimulated adenylate cyclase exhibits predominantly the benzyl alcohol-insensitive component.

scholarly journals Guanosine 5′-triphosphate and guanosine 5′-[βγ-imido]triphosphate effect a collision coupling mechanism between the glucagon receptor and catalytic unit of adenylate cyclase

1980 ◽  
Vol 186 (3) ◽  
pp. 649-658 ◽  
Author(s):  
Miles D. Houslay ◽  
Irene Dipple ◽  
Keith R. F. Elliott
Keyword(s):  
Adenylate Cyclase ◽  
Plasma Membranes ◽  
Specific Binding ◽  
Regulatory Protein ◽  
Cyclase Activity ◽  
Adenylate Cyclase Activity ◽  
Lipid Phase ◽  
Trypsin Treatment ◽  
Phase Separations ◽  
Catalytic Unit

1. GTP, but not p[NH]ppG (guanosine 5′-[βγ-imido]triphosphate), abolishes the sensitivity of glucagon-stimulated adenylate cyclase to the lipid-phase separations occurring in the outer half of the bilayer in liver plasma membranes from rat. 2. When either GTP or p[NH]ppG alone stimulate adenylate cyclase, the enzyme senses only those lipid-phase separations occurring in the inner half of the bilayer. 3. Trypsin treatment of intact hepatocytes has no effect on the basal, fluoride-, GTP- or p[NH]ppG-stimulated adenylate cyclase activity. However, 125I-labelled-glucagon specific binding decays with a half-life matching that of the decay of glucagon-stimulated adenylate cyclase activity. 4. When GTP or p[NH]ppG are added to assays of glucagon-stimulated activity, the half-life of the trypsin-mediated decay of activity is substantially increased and the decay plots are no longer first-order. 5. Trypsin treatment of purified rat liver plasma membranes abolishes basal and all ligand-stimulated adenylate cyclase activity, and 125I-labelled-glucagon specific binding. 6. Benzyl alcohol activates the GTP- and p[NH]ppG-stimulated activities in an identical fashion, whereas these activities are affected differently when glucagon is present in the assays. 7. We suggest that guanine nucleotides alter the mode of coupling between the receptor and catalytic unit. In the presence of glucagon and GTP, a complex of receptor, catalytic unit and nucleotide regulatory protein occurs as a transient intermediate, releasing a free unstable active catalytic unit. In the presence of p[NH]ppG and glucagon, the transient complex yields a relatively stable complex of the catalytic unit associated with a p[NH]ppG-bound nucleotide-regulatory protein.

scholarly journals Prostaglandin-Endoperoxides and Cyclic 3′-5′-AMP in Platelets of Patients with Uremia

1977 ◽  
Author(s):  
F. R. Matthias ◽  
W. Palinski
Keyword(s):  
Renal Failure ◽  
Chronic Renal Failure ◽  
Adenylate Cyclase ◽  
Prostaglandin E1 ◽  
Cyclase Activity ◽  
Adenylate Cyclase Activity ◽  
Bleeding Tendency ◽  
Prostaglandin Endoperoxide ◽  
Uremic Patients

In platelets of normal donors and of patients with chronic renal failure the following determinations were performed: 1. prostaglandin-endoperoxide-formation after N-ethylmaleimide stimulation measured as malondialdehyde; 2. the c-AMP level according to the Gilman-method; 3. the adenylate-cyclase activity in response to prostaglandin E1; 4. aggregation in response to collagen.In comparison to’normal donores the prostaglandin-endoperoxide production was reduced in uremic patients. Plasma of uremics depresses the endoperoxide formation of normal platelets. The basal c-AMP level of platelets of’ patients with renal failure was not significantly changed, whereas the plasma c-AMP level was increased; the activation of the platelets adenylate-cyclase was impaired. The adenylate-cyclase activity of platelets from normal donors was reduced by uremic plasma.. The results are of interest as to the explanation of the bleeding tendency of uremic patients.

Effects of benzyl alcohol on PTH receptor-adenylate cyclase system of canine kidney

1985 ◽  
Vol 248 (1) ◽  
pp. E31-E35
Author(s):  
K. J. Martin ◽  
C. L. McConkey ◽  
T. J. Stokes
Keyword(s):  
Enzyme Activity ◽  
Adenylate Cyclase ◽  
Benzyl Alcohol ◽  
Membrane Fluidity ◽  
Phospholipid Composition ◽  
Cyclase Activity ◽  
Adenylate Cyclase Activity ◽  
Adenylate Cyclase System ◽  
Hormone Sensitive ◽  
Canine Kidney

In many systems, perturbations of membrane architecture by changes of lipid and phospholipid composition have been shown to alter the activity of membrane-bound enzymes. The present studies examined the effect of benzyl alcohol, an agent that has been shown to increase membrane fluidity, on the parathyroid hormone (PTH)-sensitive adenylate cyclase system of canine kidney. Benzyl alcohol progressively increased basal adenylate cyclase activity up to fourfold and maximal enzyme activity in the presence of PTH, GTP, guanylimidodiphosphate, and sodium fluoride by four- to sixfold. In the presence of 20 mM Mn2+ (no Mg2+), conditions under which enzyme activity is devoid of influence of guanine nucleotides or hormones, benzyl alcohol was without effect. PTH binding was increased by 25% in the presence of benzyl alcohol without a change in binding affinity. Fluorescent polarization studies using diphenylhexatriene showed a decrease in fluorescence anisotropy in the presence of benzyl alcohol. The results suggest that benzyl alcohol facilitates the interaction of the components of the adenylate cyclase system, presumably by increasing membrane fluidity. Alterations of membrane fluidity may be a potent means of regulating hormone sensitive adenylate cyclase activity.

scholarly journals The activity of glucagon-stimulated adenylate cyclase from rat liver plasma membranes is modulated by the fluidity of its lipid environment

1978 ◽  
Vol 174 (1) ◽  
pp. 179-190 ◽  
Author(s):  
I Dipple ◽  
M D Houslay
Keyword(s):  
Adenylate Cyclase ◽  
Benzyl Alcohol ◽  
Plasma Membranes ◽  
Break Point ◽  
Alcohol Concentration ◽  
Cyclase Activity ◽  
Adenylate Cyclase Activity ◽  
Lipid Phase ◽  
Glucagon Receptor ◽  
Arrhenius Plots

1. The local anaesthetic benzyl alcohol progressively activated glucagon-stimulated adenylate cyclase activity up to a maximum at 50 mM-benzyl alcohol. Further increases in benzyl alcohol concentration inhibited the activity. The fluoride-stimulated adenylate cyclase activity was similarly affected except for an inhibition of activity occurring at low benzyl alcohol concentrations (approx. 10 mM. 2. The fluoride-stimulated adenylate cyclase activity of a solubilized enzyme preparation was unaffected by any of the benzyl alcohol concentrations tested. 3. Increases in 3-phenylpropan-1-ol and 5-phenylpentan-1-ol concentrations progressively activated both the fluoride- and glucagon-stimulated adenylate cyclase activities up to a maximum, above which further increases in alcohol concentration inhibited the activities. 4. The ‘break’ points in Arrhenius plots of glucagon-stimulated adenylate cyclase activity in native plasma membranes, and in plasma membranes fused with synthetic dimyristoyl phosphatidylcholine so as to constitute 60% of the total lipid pool, were decreased by approx. 6 degrees C by addition of 40 mM-benzyl alcohol. This was accompanied by a fall in the associated activation energies. 6. Arrhenius plots of fluoride-stimulated adenylate cyclase activity in the presence and absence of 40 mM-benzyl alcohol were linear, although addition of benzyl alcohol caused a dramatic decrease in the associated activation energy of the reaction. 7. 5′-Nucleotidase activity was stimulated by benzyl alcohol, and the ‘break’ point in the Arrhenius plot of its activity was decreased by about 6 degrees C by addition of 40 mM-benzyl alcohol to the assay. 8. It is suggested that benzyl alcohol effects a fluidization of the bilayer, which is clearly demonstrated by its ability to lower the temperature of a lipid phase separation occurring at 28 degrees C in the outer half of the bilayer to around 22 degrees C. The increase in bilayer fluidity relieves a physical constraint on the membrane-bound adenylate cyclase, activating the enzyme. 9. The various inhibition phenomena are discussed in detail, together with the suggestion that the interaction between the uncoupled catalytic unit of adenylate cyclase and the lipids of the bilayer is altered on its physical coupling to the glucagon receptor.

Effect of lithium on prostaglandin E1− stimulated adenylate cyclase activity of human platelets

1974 ◽  
Vol 23 (4) ◽  
pp. 845-855 ◽  
Author(s):  
Wang Yao-Chun ◽  
Ghanshyam N. Pandey ◽  
Joe Mendels ◽  
Alan Frazer
Keyword(s):  
Adenylate Cyclase ◽  
Prostaglandin E1 ◽  
Human Platelets ◽  
Cyclase Activity ◽  
Adenylate Cyclase Activity
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