Willmott N, Sethi J K, Walseth T F, Lee H C, White A M, Galione A
Department of Pharmacology, Oxford University, Oxford OX1 3QT, United Kingdom.
J Biol Chem. 1996 Feb 16;271(7):3699-705. doi: 10.1074/jbc.271.7.3699.
Cyclic adenosine diphosphate ribose (cADPR) is a potent endogenous calcium-mobilizing agent synthesized from beta-NAD+ by ADP-ribosyl cyclases in sea urchin eggs and in several mammalian cells (Galione, A., and White, A. (1994) Trends Cell Biol. 4, 431 436). Pharmacological studies suggest that cADPR is an endogenous modulator of Ca2+-induced Ca2+ release mediated by ryanodine-sensitive Ca2+ release channels. An unresolved question is whether cADPR can act as a Ca2+-mobilizing intracellular messenger. We show that exogenous application of nitric oxide (NO) mobilizes Ca2+ from intracellular stores in intact sea urchin eggs and that it releases Ca2+ and elevates cADPR levels in egg homogenates. 8-Amino-cADPR, a selective competitive antagonist of cADPR-mediated Ca2+ release, and nicotinamide, an inhibitor of ADP-ribosyl cyclase, inhibit the Ca2+-mobilizing actions of NO, while, heparin, a competitive antagonist of the inositol 1,4,5-trisphosphate receptor, did not affect NO-induced Ca2+ release. Since the Ca2+-mobilizing effects of NO can be mimicked by cGMP, are inhibited by the cGMP-dependent-protein kinase inhibitor, Rp-8-pCPT-cGMPS, and in egg homogenates show a requirement for the guanylyl cyclase substrate, GTP, we suggest a novel action of NO in mobilizing intracellular calcium from microsomal stores via a signaling pathway involving cGMP and cADPR. These results suggest that cADPR has the capacity to act as a Ca2+-mobilizing intracellular messenger.
环磷酸腺苷二磷酸核糖(cADPR)是一种强效的内源性钙动员剂,由海胆卵和多种哺乳动物细胞中的ADP - 核糖基环化酶从β - NAD +合成(加利奥内,A.,和怀特,A.(1994年)《细胞生物学趋势》4,431 - 436)。药理学研究表明,cADPR是由雷诺丁敏感的钙释放通道介导的钙诱导钙释放的内源性调节剂。一个尚未解决的问题是cADPR是否能作为一种钙动员的细胞内信使。我们发现,外源性应用一氧化氮(NO)可从完整海胆卵的细胞内储存中动员钙,并且它能在卵匀浆中释放钙并提高cADPR水平。8 - 氨基 - cADPR是cADPR介导的钙释放的选择性竞争性拮抗剂,烟酰胺是ADP - 核糖基环化酶的抑制剂,它们抑制NO的钙动员作用,而肝素是肌醇1,4,5 - 三磷酸受体的竞争性拮抗剂,不影响NO诱导的钙释放。由于NO的钙动员作用可被cGMP模拟,被cGMP依赖性蛋白激酶抑制剂Rp - 8 - pCPT - cGMPS抑制,并且在卵匀浆中显示需要鸟苷酸环化酶底物GTP,我们提出NO通过涉及cGMP和cADPR的信号通路从微粒体储存中动员细胞内钙的新作用。这些结果表明cADPR有能力作为一种钙动员的细胞内信使。