title: Studies of solute self-association by sedimentation equilibrium: allowance for effects of thermodynamic non-ideality beyond the consequences of nearest-neighbor interactions.
abstract: A sedimentation equilibrium study of alpha-chymotrypsin self-association in acetate-chloride buffer, pH 4.1 I 0.05, has been used to illustrate determination of a dimerization constant under conditions where thermodynamic non-ideality is manifested beyond the consequences of nearest-neighbor interactions. Because the expressions for the experimentally determinable interaction parameters comprise a mixture of equilibrium constant and excluded volume terms, the assignment of reasonable magnitudes to the relevant virial coefficients describing non-associative cluster formation is essential for the evaluation of a reliable estimate of the dimerization constant. Determination of these excluded volume parameters by numerical integration over the potential-of-mean-force is shown to be preferable to their calculation by approximate analytical solutions of the integral for this relatively small enzyme monomer with high net charge (+10) under conditions of low ionic strength (0.05 M).
authors.affiliation: Department of Physics, University of Auckland, Auckland, New Zealand.
authors.forename: P R
authors.initials: PR
authors.lastname: Wills
authors.forename: D J
authors.initials: DJ
authors.lastname: Winzor
mesh_terms: D000465:Algorithms; D002498:Centrifugation; D002918:Chymotrypsin / Q000737:chemistry*; D017711:Nonlinear Dynamics; D013816:Thermodynamics
chemical_list: D002918:Chymotrypsin