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J. Biol. Chem., Vol. 277, Issue 38, 35682-35687, September 20, 2002
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From the Departments of Calcium plays a regulatory role in
several aspects of protein trafficking in the cell. Both vesicle fusion
and vesicle formation can be inhibited by the addition of calcium
chelators. Because the effects of calcium chelators have been studied
predominantly in cell-free systems, it is not clear exactly which
transport steps in the secretory pathway are sensitive to calcium
levels. In this regard, we have studied the effects of calcium
chelators on both anterograde and retrograde protein transport in whole cells. Using both cytochemical and biochemical analyses, we find that
the anterograde-directed exit of vesicular stomatitis virus G
protein and the retrograde-directed exit of Shiga toxin from the Golgi
apparatus are both inhibited by calcium chelation. The exit of
vesicular stomatitis virus G from a pre-Golgi compartment and the exit
of Shiga toxin from an endosomal compartment are sensitive to the
membrane-permeant calcium chelator 1,2-bis(2-amino phenoxy)ethane-N,N,N',N'-tetraacetic
acid-tetrakis (acetoxymethyl ester) (BAPTA-AM). By contrast,
endoplasmic reticulum exit and endocytic internalization from the
plasma membrane are not affected by BAPTA. Together, our data show that
some, but not all, trafficking steps in the cell may be regulated by
calcium. These studies provide a framework for a more detailed analysis
of the role of calcium as a regulatory agent during protein transport.
Selective Effects of Calcium Chelators on Anterograde and
Retrograde Protein Transport in the Cell*
,¶
Physiology & Biophysics and
§ Internal Medicine, Roy J. and Lucille A. Carver
College of Medicine, The University of Iowa,
Iowa City, Iowa 52242
*
This work was supported by grants (to M. S.) from the
American Cancer Society, The American Heart Association Heartland
Affiliate, and the Roy J. Carver Charitable Trust.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.
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