Reducing reagent-induced activation of guanylate cyclase in the cellular slime mold, Dictyostelium discoideum.
Binding of folic acid (an intrinsic agonist) to the cell surface receptors evokes transmembrane signals for activation and adaptation of guanylate cyclase in Dictyostelium discoideum. The activation signal activates this enzyme and then the adaptation signal terminates the activation. As a result, these two signals cooperatively induce a transient activation of guanylate cyclase. We investigated transmembrane signal transduction for guanylate cyclase using 2,3-dimercapto-1-propanol (BAL, a thiol-reducing reagent) since BAL induces or modifies the transmembrane signal(s). We found that BAL induced prolonged or continuous activation of guanylate cyclase. Thus, the mode of the activation is drastically different (transient versus continuous) between folic acid and BAL. We also found that the BAL-induced continuous activation was not observed when the cells were stimulated with BAL + folic acid, while folic acid + BAL transiently induced more cGMP accumulation than folic acid alone. We lastly showed that K252a, a protein kinase inhibitor, enhanced both the folic acid-induced and the BAL-induced activation of guanylate cyclase. Our results suggest that BAL induces or mimics the activation signal for guanylate cyclase. The lack of termination in the BAL-induced activation suggests that BAL does not induce the adaptation signal or that the adaptation does not inhibit the BAL-induced activation. The former possibility is more likely since folic acid suppresses the BAL-induced continuous activation. The effect of K252a suggests that protein phosphorylation plays a role in suppression of guanylate cyclase.
Department of Botany, Faculty of Science, Kyoto University.
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Oyama
D000818:Animals; D002227:Carbazoles / Q000494:pharmacology; D006152:Cyclic GMP / Q000378:metabolism; D004023:Dictyostelium / Q000187:drug effects / Q000201:enzymology* / Q000378:metabolism; D004112:Dimercaprol / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D005492:Folic Acid / Q000494:pharmacology; D006162:Guanylate Cyclase / Q000378:metabolism*; D007202:Indicators and Reagents; D026121:Indole Alkaloids; D010084:Oxidation-Reduction; D011493:Protein Kinase C / Q000037:antagonists & inhibitors
D002227:Carbazoles; D007202:Indicators and Reagents; D026121:Indole Alkaloids; D004112:Dimercaprol; D005492:Folic Acid; C049985:staurosporine aglycone; D011493:Protein Kinase C; D006162:Guanylate Cyclase; D006152:Cyclic GMP