Thermodynamics of ligand binding to P450 2B4 and P450eryF studied by isothermal titration calorimetry.
Structural plasticity and cooperativity in ligand recognition are two key aspects of the catalytic diversity of cytochrome P450 enzymes. As more mammalian P450 crystal structures have emerged, computational modeling has become a major tool to predict drug metabolism and interactions. There is a need for real solution thermodynamic data to support modeling and crystallographic observations. Using isothermal titration calorimetry (ITC) we successfully evaluated the conformational flexibility of P450 2B4 in binding imidazole inhibitors of different size and chemistry and dissected the stoichiometry and energetics of ligand binding allostery in P450eryF. Thermodynamic signatures obtained by ITC nicely correlated with structural and modeling results. Thus, ITC is a powerful tool to study structure-function relationships in P450s.
Muralidhara
B K
BK
Department of Pharmacology and Toxicology, University of Texas Medical Branch, Galveston, Texas 77555, USA. bkmurali@utmb.edu
Halpert
James R
JR
D000818:Animals; D001189:Aryl Hydrocarbon Hydroxylases / Q000737:chemistry / Q000378:metabolism*; D002151:Calorimetry; D000072467:Cytochrome P450 Family 2; D050982:GATA1 Transcription Factor / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D008024:Ligands; D011485:Protein Binding; D013816:Thermodynamics