The self-association of zinc-free bovine insulin. A single model based on interactions in the crystal that describes the association pattern in solution at pH 2, 7 and 10.
Sedimentation equilibrium studies are used to establish that a new pattern for the self-association of zinc-free insulin in solution is applicable over a wide range of conditions of pH, ionic strength and temperature. In this pattern, which is based on information from the existing literature on the X-ray crystal structure of insulin, the insulin monomer is viewed as having two distinct faces both capable of self-interaction. Sedimentation equilibrium experiments were analysed using expressions formulated for this association pattern that describe the dependence of weight average molecular weight and monomer concentration on total protein concentration. It has thereby been possible to obtain values for the two association constants which govern the system for each set of conditions studied, due allowance having been made for composition dependent non-ideality effects. Furthermore, by relating the pH, temperature and ionic strength dependence of the association constants with properties of various amino acid residues on the surface of the insulin monomer, it has also been possible to assign tentatively each constant to a particular reaction domain.
Mark
A E
AE
John Curtin School of Medical Research, Australian National University, Canberra.
Nichol
L W
LW
Jeffrey
P D
PD
D000818:Animals; D002417:Cattle; D003460:Crystallization; D006863:Hydrogen-Ion Concentration; D007328:Insulin / Q000378:metabolism*; D046911:Macromolecular Substances; D008433:Mathematics; D008962:Models, Theoretical; D012996:Solutions; D013816:Thermodynamics; D015032:Zinc
D007328:Insulin; D046911:Macromolecular Substances; D012996:Solutions; D015032:Zinc