Sucrose density gradient centrifugation of human myometrial and myomal progestin receptors

Abstract

Progestin receptors present in cytosols of myomal and myometrial origin were analyzed on sucrose density gradients of low ionic strength using a vertical tube rotor. Short term incubations (90 min) contained mainly 8S receptors. Most myometrial but few myomal preparations contained an additional 4S component, the presence of which was hormone dependent. None could be detected if the serum concentration of estradiol was high. Myometrial receptors incubated over night exhibited only a 4S peak, whereas many myomal ones still sedimented as 8S peaks. The 8 to 4S shift represented a transformation of the receptors and not a sequential interaction of the hormone with first an 8S and then a 4S entity. The transforming activity contained in the myometrial cytosol could also attack the myomal receptor as shown by mixing experiments. Only the 8S peak was observed, whenever the cytosol was first fractionated and then incubated with the hormone. Thus the transformation occurred only in presence of [ 3H]promegestone. It could be suppressed with protease inhibitors. It is concluded that these tissues contain a receptor processing protease which (1) is much less prominent in myomal than myometrial cytosol, which (2) attacks only the occupied receptors, and (3) the activity of which is hormone dependent.