tailieunhanh - Báo cáo khoa học: Inhibitor-mediated stabilization of the conformational structure of a histone deacetylase-like amidohydrolase

Histone deacetylases are major regulators of eukaryotic gene expression. Not unexpectedly, histone deacetylases are among the most promising tar-gets in cancer therapy. However, despite huge efforts in histone deacetylase inhibitor design, very little is known about the impact of histone deacety-lase inhibitors on enzyme stability. | ễFEBS Journal Inhibitor-mediated stabilization of the conformational structure of a histone deacetylase-like amidohydrolase Stefanie Kern1 Daniel Riester2 Christian Hildmann2 Andreas Schwienhorst2 and Franz-Josef Meyer-Almes1 1 Department of ChemicalEngineering and Biotechnology Darmstadt University of Applied Sciences Germany 2 Institut fur Molekulare Genetik und Praeparative Molekularbiologie Institut fuer Mikrobiologie und Genetik Goettingen Germany Keywords FB188 HDAH histone deacetylase protein denaturation protein stablization Correspondence . Meyer-Almes Department of Chemical Engineering and Biotechnology University of Applied Sciences Darmstadt Schnittspahnstr. 12 64287 Darmstadt Germany Fax 1649 6151168404 Tel 1649 6151168406 E-mail meyer-almes@ Website http cub Received 24 January 2007 revised 11 May 2007 accepted 15 May 2007 doi Histone deacetylases are major regulators of eukaryotic gene expression. Not unexpectedly histone deacetylases are among the most promising targets in cancer therapy. However despite huge efforts in histone deacetylase inhibitor design very little is known about the impact of histone deacetylase inhibitors on enzyme stability. In this study the conformational stability of a well-established histone deacetylase homolog with high structural similarity histone deacetylase-like amidohydrolase from Bordetella Alcali-genes species FB188 was investigated using denaturation titrations and stopped-flow kinetics. Based on the results of these complementary approaches we conclude that the interconversion of native histone deacetylase-like amidohydrolase into its denatured form involves several intermediates possessing different enzyme activities and conformational structures. The refolding kinetics has shown to be strongly dependent on Zn2 and to a lesser extent on K which underlines their importance not only for catalytic function but also for maintaining the correct conformational .