Development and optimization of a binding assay for histone deacetylase 4 using surface plasmon resonance.
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.
Mattu
Marco
M
Istituto di Ricerche di Biologia Molecolare P. Angeletti, Via Pontina Km 30.600, 00040 Pomezia (Rome), Italy. marco_mattu@merck.com
Di Giovine
Paolo
P
Steinkuhler
Christian
C
De Francesco
Raffaele
R
Altamura
Sergio
S
Cecchetti
Ottavia
O
Carfì
Andrea
A
Barbato
Gaetano
G
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
D004791:Enzyme Inhibitors; D004800:Enzymes, Immobilized; D056572:Histone Deacetylase Inhibitors; D007659:Ketones; D011994:Recombinant Proteins; D006655:Histone Deacetylases