Comparative binding character of two general anaesthetics for sites on human serum albumin

• DOI: 10.1042/bj20031652
• Publication Date: 2004-06-17T09:17:42Z
• Authors: Renyu LIU; Qingcheng MENG; Jin XI; Jinsheng YANG; Chung-Eun HA; Nadhipuram V. BHA...; Roderic G. ECKENHOFF
• Keywords: Binding Sites; Anesthetics, General; Static Electricity; Hydrogen Bonding; Calorimetry; Tritium; Peptide Fragments; Protein Structure, Tertiary; 03 medical and health sciences; 0302 clinical medicine; Humans; Halothane; Hydrophobic and Hydrophilic Interactions; Propofol; Serum Albumin; Chromatography, Liquid; Hydrogen; Protein Binding

Propofol and halothane are clinically used general anaesthetics, which are transported primarily by HSA (human serum albumin) in the blood. Binding characteristics are therefore of interest for both the pharmacokinetics and pharmacodynamics of these drugs. We characterized anaesthetic–HSA interactions in solution using elution chromatography, ITC (isothermal titration calorimetry), hydrogen-exchange experiments and geometric analyses of high-resolution structures. Binding affinity of propofol to HSA was determined to have a Kd of 65 µM and a stoichiometry of approx. 2, whereas the binding of halothane to HSA showed a Kd of 1.6 mM and a stoichiometry of approx. 7. Anaesthetic–HSA interactions are exothermic, with propofol having a larger negative enthalpy change relative to halothane. Hydrogen-exchange studies in isolated recombinant domains of HSA showed that propofol-binding sites are primarily found in domain III, whereas halothane sites are more widely distributed. Both location and stoichiometry from these solution studies agree with data derived from X-ray crystal-structure studies, and further analyses of the architecture of sites from these structures suggested that greater hydrophobic contacts, van der Waals interactions and hydrogen-bond formation account for the stronger binding of propofol as compared with the less potent anaesthetic, halothane.