Purification of a cortisol binding protein from hepatic plasma membrane

Abstract

A cortisol binding protein from rat liver plasma membranes has been solubilized in active form by using the zwitterionic detergent CHAPS. Two types of binding sites have been characterised in both native and solubilized membranes. The first is of high affinity and low binding capacity (12 nM; 946 fmol/mg) and the other one is of low affinity and high capacity of binding (344 nM; 12 677 fmol/mg) for solubilized membranes. The purified material retained a binding activity comparable to that displayed by the original membrane. The specific binding activity was enriched about 12700-fold, with an 8% yield. Analysis of the purified preparation on sodium dodecyl sulphate-polyacrylamide gel electrophoresis showed two protein subunits with molecular mass of 52 000 and 57 000 Da. The new cortisol-specific binding membrane protein could be related to the nongenomic effects previously described for this hormone.