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Originally published In Press as doi:10.1074/jbc.M005316200 on June 27, 2000

J. Biol. Chem., Vol. 275, Issue 38, 29233-29237, September 22, 2000
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Regulation of Intra-Golgi Membrane Transport by Calcium*

Amir Porat and Zvulun ElazarDagger

From the Department of Biological Chemistry, The Weizmann Institute of Science, Rehovot 76100, Israel

Calcium cations play a critical role in regulating vesicular transport between different intracellular membrane-bound compartments. The role of calcium in transport between the Golgi cisternae, however, remains unclear. Using a well characterized cell-free intra-Golgi transport assay, we now show that changes in free Ca2+ concentration in the physiological range regulate this transport process. The calcium-chelating agent 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid blocked transport with an IC50 of approximately 0.8 mM. The effect of 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid was reversible by addition of fresh cytosol and was irreversible when performed in the presence of a Ca2+ ionophore that depletes calcium from lumenal stores. We demonstrate here that intra-Golgi transport is stimulated by low Ca2+ concentrations (20-100 nM) but is inhibited by higher concentrations (above 100 nM). Further, we show that calmodulin antagonists specifically block intra-Golgi transport, implying a role for calmodulin in mediating the effect of calcium. Our results suggest that Ca2+ efflux from intracellular pools may play an essential role in regulating intra-Golgi transport.

* This work was supported in part by the Israel Science Foundation.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 Incumbent of the Sholimo and Michla Tomarin Career Development Chair of Membrane Physiology. To whom correspondence should be addressed: Tel.: 972-8-9343682; Fax: 972-8-9344112; E-mail: bmzevi@wicc.weizmann.ac.il

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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