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J. Biol. Chem., Vol. 280, Issue 17, 16882-16890, April 29, 2005

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The Intermediate Filament Protein Vimentin Is a New Target for Epigallocatechin Gallate^*

Svetlana Ermakova, Bu Young Choi, Hong Seok Choi, Bong Seok Kang, Ann M. Bode, and Zigang Dong

From the Hormel Institute, University of Minnesota, Austin, Minnesota 55912

Epigallocatechin gallate (EGCG) is the major active polyphenol in green tea. Protein interaction with EGCG is a critical step in the effects of EGCG on the regulation of various key proteins involved in signal transduction. We have identified a novel molecular target of EGCG using affinity chromatography, two-dimensional electrophoresis, and mass spectrometry for protein identification. Spots of interest were identified as the intermediate filament, vimentin. The identification was confirmed by Western blot analysis using an anti-vimentin antibody. Experiments using a pull-down assay with [³H]EGCG demonstrate binding of EGCG to vimentin with a K_d of 3.3 nM. EGCG inhibited phosphorylation of vimentin at serines 50 and 55 and phosphorylation of vimentin by cyclin-dependent kinase 2 and cAMP-dependent protein kinase. EGCG specifically inhibits cell proliferation by binding to vimentin. Because vimentin is important for maintaining cellular functions and is essential in maintaining the structure and mechanical integration of the cellular space, the inhibitory effect of EGCG on vimentin may further explain its anti-tumor-promoting effect.

Received for publication, December 17, 2004 , and in revised form, February 8, 2005.

^* This work was supported in part by the Hormel Foundation and National Institutes of Health Grants CA81064 and CA88961. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed: Hormel Institute, University of Minnesota, 801 16th Ave. NE, Austin, MN 55912. Tel.: 507-437-9600; Fax: 507-437-9606; E-mail: zgdong{at}hi.umn.edu.

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