Reversible dissociation of chick oviduct progesterone receptor subunits.

We report here the reversible dissociation of chick oviduct progesterone receptor subunits by pyridoxal 5'-phosphate. This agent has been reported to inhibit binding of steroid hormone receptors to DNA and nuclei (Cake et al., 1978). We have found that pyridoxal 5'-phosphate inhibits binding of chick oviduct progesterone receptors to DNA-cellulose, and also dissociates 6-8S cytosolic receptor aggregates to 4S monomers. Both of these effects are shown to be reversible if pyridoxal phosphate is removed, allowing in vitro reconstitution of receptor aggregates. Fidelity of reconstitution has been assessed by testing the reconstituted aggregate for binding to DNA-cellulose, phosphocellulose, and by studies using sedimentation velocity measurements. By these three criteria, the reconstituted product is indistinguishable from the native cytosol complex from which the monomers were derived. The reconstitution reaction shows an absolute requirement for the presence of both receptor monomers A and B. Titration experiments show a molar ratio of 1:1 for A and B in the reconstituted aggregates. These reconstitution studies confirm our hypothesis (originally based upon dissociation experiments) that native receptor aggregates are composed of the A and B proteins as subunits.