Lübker Carolin, Seifert Roland
Institute of Pharmacology, Hannover Medical School, Hannover, Germany.
PLoS One. 2015 May 6;10(5):e0124017. doi: 10.1371/journal.pone.0124017. eCollection 2015.
Adenylyl cyclases (ACs) catalyze the conversion of ATP into the second messenger cAMP. Membranous AC1 (AC1) is involved in processes of memory and learning and in muscle pain. The AC toxin edema factor (EF) of Bacillus anthracis is involved in the development of anthrax. Both ACs are stimulated by the eukaryotic Ca(2+)-sensor calmodulin (CaM). The CaM-AC interaction could constitute a potential target to enhance or impair the AC activity of AC1 and EF to intervene in above (patho)physiological mechanisms. Thus, we analyzed the impact of 39 compounds including typical CaM-inhibitors, an anticonvulsant, an anticholinergic, antidepressants, antipsychotics and Ca(2+)-antagonists on CaM-stimulated catalytic activity of AC1 and EF. Compounds were tested at 10 μM, i.e., a concentration that can be reached therapeutically for certain antidepressants and antipsychotics. Calmidazolium chloride decreased CaM-stimulated AC1 activity moderately by about 30%. In contrast, CaM-stimulated EF activity was abrogated by calmidazolium chloride and additionally decreased by chlorpromazine, felodipine, penfluridol and trifluoperazine by about 20-40%. The activity of both ACs was decreased by calmidazolium chloride in the presence and absence of CaM. Thus, CaM-stimulated AC1 activity is more insensitive to inhibition by small molecules than CaM-stimulated EF activity. Inhibition of AC1 and EF by calmidazolium chloride is largely mediated via a CaM-independent allosteric mechanism.
腺苷酸环化酶(ACs)催化ATP转化为第二信使环磷酸腺苷(cAMP)。膜性AC1参与记忆和学习过程以及肌肉疼痛。炭疽芽孢杆菌的AC毒素水肿因子(EF)参与炭疽病的发展。两种ACs均受真核生物钙传感器钙调蛋白(CaM)的刺激。CaM与AC的相互作用可能构成一个潜在靶点,用于增强或削弱AC1和EF的AC活性,从而干预上述(病理)生理机制。因此,我们分析了39种化合物的影响,这些化合物包括典型的CaM抑制剂、一种抗惊厥药、一种抗胆碱能药、抗抑郁药、抗精神病药和钙拮抗剂,它们对CaM刺激的AC1和EF的催化活性的影响。化合物在10 μM浓度下进行测试,即某些抗抑郁药和抗精神病药在治疗时可达到的浓度。氯化氯米帕明使CaM刺激的AC1活性适度降低约30%。相比之下,CaM刺激的EF活性被氯化氯米帕明消除,并且氯丙嗪、非洛地平、五氟利多和三氟拉嗪使其额外降低约20%-40%。在有和没有CaM的情况下,氯化氯米帕明均降低了两种ACs的活性。因此,CaM刺激的AC1活性比CaM刺激的EF活性对小分子抑制更不敏感。氯化氯米帕明对AC1和EF的抑制很大程度上是通过一种不依赖CaM的变构机制介导的。