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Originally published In Press as doi:10.1074/jbc.M110944200 on February 20, 2002

J. Biol. Chem., Vol. 277, Issue 19, 17255-17262, May 10, 2002
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Dichotomy of Ca2+ Signals Triggered by Different Phospholipid Pathways in Antigen Stimulation of Human Mast Cells*

Alirio J. MelendezDagger and Aik Kia Khaw

From the Department of Physiology, Faculty of Medicine, National University of Singapore, Singapore 117597, Singapore

Mast cell activation triggers Ca2+ signals and the release of enzyme-containing granules, events that play a major role in allergic/hypersensitivity reactions. However, the precise molecular mechanisms that regulate antigen-triggered degranulation and Ca2+ fluxes in human mast cells are still poorly understood. Here we show, for the first time, that a receptor can trigger Ca2+ via two separate molecular mechanisms. Using an antisense approach, we show that IgE-antigen stimulation of human bone marrow-derived mast cells triggers a sphingosine kinase (SPHK) 1-mediated fast and transient Ca2+ release from intracellular stores. However, phospholipase C (PLC) gamma 1 triggers a second (slower) wave of calcium release from intracellular stores, and it is this PLCgamma 1-generated signal that is responsible for Ca2+ entry. Surprisingly, Fcepsilon RI (a high affinity receptor for IgE)-triggered mast cell degranulation depends on the first, sphingosine kinase-mediated Ca2+ signal. These two pathways act independently because antisense knock down of either enzyme does not interfere with the activity of the other enzyme. Of interest, similar to PLCgamma 1, SPHK1 translocates rapidly to the membrane after Fcepsilon RI cross-linking. Here we also show that SPHK1 activity depends on phospholipase D1 and that Fcepsilon RI-triggered mast cell degranulation depends primarily on the activation of both phospholipase D1 and SPHK1.

* This work was supported by a start-up grant from the National Medical Research Counsel of Singapore.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger To whom correspondence should be addressed: Dept. of Physiology, Faculty of Medicine, National University of Singapore, 2 Medical Dr., MD 9, 03-04, Singapore 117597, Singapore. Tel.: 65-874-1697; Fax: 65-778-8161; E-mail: phsmraj@nus.edu.sg.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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