title: A membrane protein associated with the prolactin receptor. Studies with a photoactivatable human growth hormone derivative.
abstract: Prolactin receptor from rat liver (PRL-R, 42 kDa) was cross-linked to a radiolabeled azidophenacyl derivative of human growth hormone ([125I]AP-hGH) to yield a 63 kDa adduct. In addition, a protein of Mr 50-52 K was detected as a 73 kDa complex. Microsomes incubated with either (a) increasing amounts of [125I]AP-hGH, or (b) a fixed amount of photoprobe and increasing concentrations of unlabeled hGH, showed that the 73/63 kDa band intensity ratio remains constant (0.71-0.77). Once transferred onto nitrocellulose membranes, only the 42 kDa protein is able to bind [125I]AP-hGH or [125I]hGH. Two anti-PRL-R monoclonal antibodies fail to cross-react with proteins of Mr 50-52 K. In membranes solubilized with 3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate (CHAPS), a significantly lower amount of the 73 kDa complex is detected. Thus, the 50-52 kDa protein appears to be structurally unrelated to, but is presumably associated with the PRL-R. The 73 kDa complex is also detected under low membrane fluidity conditions (1 degree C), indicating that PRL-R associates to this 50-52 kDa protein prior to hormone binding. Perfusion of rat liver with [125I]AP-hGH shows that this associated protein accompanies the receptor along its intracellular pathway.
authors.affiliation: Instituto de Química y Fisicoquímica Biológicas, Universidad de Buenos Aires-CONICET, Argentina.
authors.forename: N T
authors.initials: NT
authors.lastname: Masckauchán
authors.forename: J M
authors.initials: JM
authors.lastname: Delfino
authors.forename: H N
authors.initials: HN
authors.lastname: Fernández
mesh_terms: D000345:Affinity Labels / Q000378:metabolism* / Q000493:pharmacokinetics; D000818:Animals; D000911:Antibodies, Monoclonal; D001386:Azides / Q000378:metabolism* / Q000493:pharmacokinetics; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D002793:Cholic Acids; D019382:Human Growth Hormone / Q000031:analogs & derivatives* / Q000378:metabolism* / Q000493:pharmacokinetics; D007457:Iodine Radioisotopes; D008099:Liver / Q000378:metabolism; D008565:Membrane Proteins / Q000737:chemistry* / Q000378:metabolism*; D010777:Photochemistry; D051381:Rats; D017207:Rats, Sprague-Dawley; D011981:Receptors, Prolactin / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D000345:Affinity Labels; D000911:Antibodies, Monoclonal; D001386:Azides; D002793:Cholic Acids; D007457:Iodine Radioisotopes; D008565:Membrane Proteins; D011981:Receptors, Prolactin; D019382:Human Growth Hormone; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate