Nitric Oxide-induced Mobilization of Intracellular Calcium via the Cyclic ADP-ribose Signaling Pathway

doi:10.1074/jbc.271.7.3699

Nitric Oxide-induced Mobilization of Intracellular Calcium via the Cyclic ADP-ribose Signaling Pathway (*)

From the (1)Department of Pharmacology, Oxford University, Oxford OX1 3QT, United Kingdom and the Departments of Pharmacology and
(2)Physiology, University of Minnesota, Minneapolis, Minnesota 55455

^¶To whom correspondence should be addressed. Tel.: 44-1865-271633; Fax: 44-1865-271853.

Abstract

Cyclic adenosine diphosphate ribose (cADPR) is a potent endogenous calcium-mobilizing agent synthesized from β-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 Ca-induced Ca release mediated by ryanodine-sensitive Ca release channels. An unresolved question is whether cADPR can act as a Ca-mobilizing intracellular messenger. We show that exogenous application of nitric oxide (NO) mobilizes Ca from intracellular stores in intact sea urchin eggs and that it releases Ca and elevates cADPR levels in egg homogenates. 8-Amino-cADPR, a selective competitive antagonist of cADPR-mediated Ca release, and nicotinamide, an inhibitor of ADP-ribosyl cyclase, inhibit the Ca-mobilizing actions of NO, while, heparin, a competitive antagonist of the inositol 1,4,5-trisphosphate receptor, did not affect NO-induced Ca release. Since the Ca-mobilizing effects of NO can be mimicked by cGMP, are inhibited by the cGMP-dependent-protein kinase inhibitor, R-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 Ca-mobilizing intracellular messenger.

Footnotes

↵^§ These authors have made an equal contribution to this report.
↵* This work was supported by grants from the Medical Research Council, Wellcome Trust (to A. G.), and National Institutes of Health (to H. C. L.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore by hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
↵¹ The abbreviations used are:

NO

nitric oxide

[Ca]

intracellular free calcium concentration

CICR

calcium-induced calcium release

cADPR

cyclic ADP-ribose

IM

intracellular medium

IP

inositol 1,4,5-trisphosphate

R-8-pCPT-cGMPS

R isomer of 8-(4-chlorophenylthio)-guanosine-3′,5′-cyclic monophosphorothioate

RyR

ryanodine receptor

Pipes

1,4-piperazinediethanesulfonic acid.
↵²J. Sethi and A. Galione, unpublished observations.
- Received October 31, 1995.
- Revision received December 7, 1995.