Preferential interaction of [35S]cysteamine with pituitary secretory granule storage forms of prolactin.
Cysteamine (CySH) inhibits the immunodetectability and bioactivity of prolactin (PRL), and we have proposed that it may act by impeding the conversion from secretory granule hormone storage forms to releasable and assayable hormone. This process appears to be dependent upon thiol:disulfide interchange reactions, which can be inhibited by the aminothiol. The present studies utilized [35S]CySH to determine whether preferential interactions could be demonstrated between CySH and bovine pituitary storage hormone forms as opposed to monomeric PRL. [35S]CySH was incubated with purified intact secretory granules, with granule 'core' preparations enriched in oligomeric forms by prior hypotonic exposure, with chromatographically isolated oligomers, and with monomeric PRL. Binding to granules was saturable and pH-dependent with greatest binding observed at pH 7.5-8.0. Binding to monomer was much less than binding to all other fractions, being 20% or less than that to any other form. HPLC studies of granules treated with [35S]CySH indicated that exposure to CySH was associated with a predominance of very high molecular weight oligomers. These forms were entrapped on the gel permeation columns, resulting in decreased protein and PRL recovery; as little as 6.6% of the PRL was eluted after 60 min of CySH exposure. CySH not only bound to storage forms of PRL but also to secretory granule membranes; whether the bioeffect is mediated through membrane modifications is unknown. Despite its relative ineffectiveness at altering growth hormone immunoactivity or secretion, CySH nonetheless also bound to growth hormone.(ABSTRACT TRUNCATED AT 250 WORDS)
Lorenson
M Y
MY
Department of Medicine, University of Rochester School of Medicine and Dentistry, N.Y.
Jacobs
L S
LS
D000818:Animals; D002417:Cattle; D002462:Cell Membrane / Q000378:metabolism; D003543:Cysteamine / Q000378:metabolism*; D003594:Cytoplasmic Granules / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D010902:Pituitary Gland / Q000166:cytology / Q000378:metabolism*; D011388:Prolactin / Q000378:metabolism*; D013347:Subcellular Fractions / Q000378:metabolism
D003543:Cysteamine; D011388:Prolactin