Histone deacetylases (HDACs) - an enzyme family that deacetylates histones and non-histone proteins - are implicated in human diseases such as cancer, and the first-generation of HDAC inhibitors are now in clinical trials. Here, we report the 2.0 Å Gresolution crystal structure of a catalytically inactive HDAC8 active-site mutant, Tyr306Phe, bound to an acetylated peptidic substrate. The structure clarifies the role of active-site residues in the deacetylation reaction and substrate recognition. Notably, the structure shows the unexpected role of a conserved residue at the active-site rim, Asp 101, in positioning the substrate by directly interacting with the peptidic backbone and imposing a constrained cis-conformation. A similar interaction is observed in a new hydroxamate inhibitor-HDAC8 structure that we also solved. The crucial role of Asp 101 in substrate and inhibitor recognition was confirmed by activity and binding assays of wild-type HDAC8 and Asp101Ala, Tyr306Phe and Asp101Ala/Tyr306Phe mutants.
Substrate binding to histone deacetylases as shown by the crystal structure of the HDAC8-substrate complex / A. Vannini, C. Volpari, P. Gallinari, P. Jones, M. Mattu, A. Carfí, R. De Francesco, C. Steinkühler, S. Di Marco. - In: EMBO REPORTS. - ISSN 1469-221X. - 8:9(2007), pp. 879-884.
|Titolo:||Substrate binding to histone deacetylases as shown by the crystal structure of the HDAC8-substrate complex|
|Parole Chiave:||HDAC; acetylated substrate; mutagenesis; crystallography; drug design|
|Settore Scientifico Disciplinare:||Settore BIO/11 - Biologia Molecolare|
|Data di pubblicazione:||2007|
|Digital Object Identifier (DOI):||http://dx.doi.org/10.1038/sj.embor.7401047|
|Appare nelle tipologie:||01 - Articolo su periodico|