Phosphorylation of bovine prolactin eliminates luteotropic activity in the rat

Phosphorylated and nonphosphorylated prolactins were isolated from bovine pituitaries and their luteotropic activity determined in female rats. Three groups of rats in day 1 of diestrus were treated i.p. twice daily for three days with 0.25 mg of either prolactin preparation or vehicle. Rats were sacrificed each day of treatment. Serum progesterone concentrations of the groups receiving vehicle or phosphorylated prolactin were similar and the vaginal cytology of these animals indicated that phosphorylated bovine prolactin (bPRL) treatment had not prolonged diestrus. Treatment with nonphosphorylated bPRL significantly increased serum progesterone concentration and the vaginal cytology indicated a diestrus prolonged for up to 4 days. Nonphosphorylated and phosphorylated bPRLs were cleared from the blood at similar rates after i.p. injection. In vitro receptor binding studies demonstrated that phosphorylated bPRL did not bind the ovarian prolactin receptor. Nonphosphorylated, but not phosphorylated, bPRL competed with radiolabeled bovine hormone for occupancy of rat ovarian prolactin receptors. These data are the first to test the activities of phosphorylated bPRL in vivo and indicate; 1) nonphosphorylated bPRL is luteotropic, 2) phosphorylated bPRL is neither luteotropic nor a prolactin receptor agonist or antagonist and 3) phosphorylated bPRL is not dephosphorylated in vivo rapidly enough to provide sufficient biologically-active bPRL to maintain luteal function.