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© The Rockefeller University Press, 0021-9525/1998//601 $5.00
The Journal of Cell Biology, Volume 141, Number 3, , 1998 601-610


Articles

Targeting of Protein Kinase C{alpha} to Caveolae



Chieko Mineo*, Yun-Shu Ying*, Christine Chapline{ddagger}, Susan Jaken{ddagger}, and Richard G.W. Anderson*

* Department of Cell Biology and Neuroscience, University of Texas Southwestern Medical Center, Dallas, Texas 75235-9039; and {ddagger} Adirondack Biomedical Research Institute, Lake Placid, New York 12946

Previously, we showed caveolae contain a population of protein kinase C{alpha} (PKC{alpha}) that appears to regulate membrane invagination. We now report that multiple PKC isoenzymes are enriched in caveolae of unstimulated fibroblasts. To understand the mechanism of PKC targeting, we prepared caveolae lacking PKC{alpha} and measured the interaction of recombinant PKC{alpha} with these membranes. PKC{alpha} bound with high affinity and specificity to caveolae membranes. Binding was calcium dependent, did not require the addition of factors that activate the enzyme, and involved the regulatory domain of the molecule. A 68-kD PKC{alpha}-binding protein identified as sdr (serum deprivation response) was isolated by interaction cloning and localized to caveolae. Antibodies against sdr inhibited PKC{alpha} binding. A 100–amino acid sequence from the middle of sdr competitively blocked PKC{alpha} binding while flanking sequences were inactive. Caveolae appear to be a membrane site where PKC enzymes are organized to carry out essential regulatory functions as well as to modulate signal transduction at the cell surface.


Abbreviations used in this paper: DAG, diacylglycerol; MBP, maltose-binding protein; PKC, protein kinase C; PMA, phorbol-12-myristate-13-acetate; PS, phosphatidylserine; sdr, serum deprivation response; RD, regulatory domain.

Address all correspondence to Richard G.W. Anderson, Department of Cell Biology and Neuroscience, University of Texas Southwestern Medical Center, Dallas, TX 75235-9039. Tel.: (214) 648-2346. Fax: (214) 648-7577. E-mail: anders06{at}utsw.swmed.edu



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