title: Development and optimization of a binding assay for histone deacetylase 4 using surface plasmon resonance.
abstract: Histone deacetylase 4 (HDAC4) is a histone deacetylase profoundly involved in cell differentiation and in the pathogenesis of cancer. The histone deacetylase inhibitors are a new, promising class of anticancer agents. The screening of molecular interactions involving determination of the affinity of drug candidates is an integral part of the drug discovery process. Here we report the development of an assay using surface plasmon resonance for the analysis of HDAC4-small molecule interactions. We describe a new cloning and purification strategy that can be used to set up surface plasmon resonance experiments with other recombinant proteins.
authors.affiliation: Istituto di Ricerche di Biologia Molecolare P. Angeletti, Via Pontina Km 30.600, 00040 Pomezia (Rome), Italy. marco_mattu@merck.com
authors.forename: Marco
authors.initials: M
authors.lastname: Mattu
authors.forename: Paolo
authors.initials: P
authors.lastname: Di Giovine
authors.forename: Christian
authors.initials: C
authors.lastname: Steinkuhler
authors.forename: Raffaele
authors.initials: R
authors.lastname: De Francesco
authors.forename: Sergio
authors.initials: S
authors.lastname: Altamura
authors.forename: Ottavia
authors.initials: O
authors.lastname: Cecchetti
authors.forename: Andrea
authors.initials: A
authors.lastname: Carfì
authors.forename: Gaetano
authors.initials: G
authors.lastname: Barbato
mesh_terms: D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000378:metabolism / Q000494:pharmacology; D004800:Enzymes, Immobilized / Q000037:antagonists & inhibitors / Q000378:metabolism; D056572:Histone Deacetylase Inhibitors; D006655:Histone Deacetylases / Q000378:metabolism*; D007659:Ketones / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D020349:Surface Plasmon Resonance
chemical_list: D004791:Enzyme Inhibitors; D004800:Enzymes, Immobilized; D056572:Histone Deacetylase Inhibitors; D007659:Ketones; D011994:Recombinant Proteins; D006655:Histone Deacetylases