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Originally published In Press as doi:10.1074/jbc.M211746200 on March 6, 2003

J. Biol. Chem., Vol. 278, Issue 21, 19199-19208, May 23, 2003
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Mechanisms of VE-cadherin Processing and Degradation in Microvascular Endothelial Cells*

Kanyan Xiao {ddagger} § ¶, David F. Allison {ddagger} § ¶, Margaret D. Kottke {ddagger} § ¶, Susan Summers {ddagger} § ¶, George P. Sorescu {ddagger} § ¶, Victor Faundez § and Andrew P. Kowalczyk {ddagger} § ¶ ||

From the {ddagger} Department of Dermatology, Emory University School of Medicine, Atlanta, Georgia 30322, § Department of Cell Biology, Emory University School of Medicine, Atlanta, Georgia 30322, Emory Skin Diseases Research Center, Emory University School of Medicine, Atlanta, Georgia 30322

VE-cadherin is an endothelial-specific cadherin that plays important roles in vascular morphogenesis and growth control. To investigate the mechanisms by which endothelial cells regulate cadherin cell surface levels, a VE-cadherin mutant containing the non-adhesive interleukin-2 (IL-2) receptor extracellular domain and the VE-cadherin cytoplasmic tail (IL-2R-VE-cadcyto) was expressed in microvascular endothelial cells. Expression of the IL-2R-VE-cadcyto mutant resulted in the internalization of endogenous VE-cadherin and in a dramatic decrease in endogenous VE-cadherin levels. The internalized VE-cadherin co-localized with early endosomes, and the lysosomal inhibitor chloroquine dramatically inhibited the down-regulation of VE-cadherin in cells expressing the IL-2R-VE-cadcyto mutant. Chloroquine treatment also resulted in the accumulation of a VE-cadherin fragment lacking the {beta}-catenin binding domain of the VE-cadherin cytoplasmic tail. The formation of the VE-cadherin fragment could be prevented by treating endothelial cells with proteasome inhibitors. Furthermore, inhibition of the proteasome prevented VE-cadherin internalization and inhibited the disruption of endothelial intercellular junctions by the IL-2RVE-cadcyto mutant. These results provide new insights into the mechanisms of VE-cadherin processing and degradation in microvascular endothelial cells.

Received for publication, November 18, 2002 , and in revised form, February 21, 2003.

|| To whom correspondence should be addressed: Dept. of Dermatology, Emory University School of Medicine, Woodruff Memorial Bldg., Rm. 5007, Atlanta, GA 30322. Tel.: 404-727-8517; Fax: 404-727-5878; E-mail: akowalc{at}emory.edu.


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