Cysteamine causes reduction of prolactin monomers followed by aggregation in the rat pituitary gland.
Storage forms of PRL were studied in control and cysteamine-treated cultures of estradiol-induced tumors in Fischer 344 rats and in secretory granules isolated from these tumors to further investigate the mechanism of action of cysteamine on PRL. The two major bands visible when protein is stained after electrophoresis of isolated granules migrate to the position of PRL and GH monomers. Electrophoresis under reducing conditions changes the position, but does not noticeably increase the amount of each band. [3H]PRL in cells labeled for 8 h with [3H]leucine also exists predominantly as monomer. Immunoreactivity of PRL in cell lysates or isolated granules is not affected by incubation with reducing agents beta-mercaptoethanol or glutathione at concentrations up to 5 mM, but cysteamine decreases PRL immunoreactivity in isolated granules at concentrations of 3 mM and higher. Electrophoresis of isolated granules after incubation with 25 mM cysteamine for 1 h demonstrates that cysteamine converts PRL to the reduced form. After 4 h, or after dilution of the granules before solubilization, the amount of reduced monomer is decreased, and larger molecular weight species appear. The reduced monomer can be recovered by electrophoresis under reducing conditions. The fully immunoreactive form can be recovered by incubation for 1 h with dithiothreitol at concentrations of 0.3 mM-3 mM. These data indicate that: PRL exists predominantly in monomeric form in the rat pituitary gland, and cysteamine reduces PRL, and formation of disulfide-linked aggregates of PRL occurs subsequently under some conditions.
D000818:Animals; D003543:Cysteamine / Q000494:pharmacology*; D003594:Cytoplasmic Granules / Q000032:analysis; D004229:Dithiothreitol / Q000494:pharmacology; D005260:Female; D013006:Growth Hormone / Q000032:analysis; D010902:Pituitary Gland / Q000032:analysis / Q000187:drug effects*; D010911:Pituitary Neoplasms / Q000032:analysis; D011388:Prolactin / Q000032:analysis*; D051381:Rats; D011916:Rats, Inbred F344; D013439:Sulfhydryl Reagents / Q000494:pharmacology
D013439:Sulfhydryl Reagents; D003543:Cysteamine; D011388:Prolactin; D013006:Growth Hormone; D004229:Dithiothreitol