Proteins associated with somatogenic and lactogenic receptors in microsomal membranes and intact rat hepatocytes.
Lactogenic and somatogenic receptors present in rat liver have been examined by cross-linking with a derivative of human somatotropin (AP-hGH1) followed by SDS-polyacrylamide gel electrophoresis. AP-hGH1, which has a content of 2.2 azidophenacyl groups per molecule, mainly linked to half Cys-182 and half Cys-189, exerted a specificity similar to that of the native hormone (hGH), with an ability of 46% with respect to hGH to compete with the radiolabelled hormone for the binding sites of microsomal preparations. Photolysis of the 125I-labelled derivative bound to the lactogenic receptors present in either microsomal membranes or Triton X-100 solubilized preparations gave rise to a 63 kDa species. In addition, 30% of the covalent complexes formed in microsomal membranes belonged to a species with a molecular mass of 70 kDa. Incubation of viable rat hepatocytes with the radiolabelled derivative at either 0 degrees C for 3 h or 15 degrees C for 1.5 h and subjection to irradiation, yielded covalent complexes of molecular masses estimated at 130, 73, 63, 45 and 35 kDa. Experiments performed in the presence of 1 mM NaCN, gave rise to the previous species in a similar yield as that obtained in the absence of cyanide. The 130 kDa complex is related to the somatogenic binding sites, since it was not visualized in the presence of unlabelled bovine somatotropin, while the 70-73, 63, 45 and 35 kDa bands disappeared when the incubations were performed in the presence of unlabelled ovine prolactin.
Robetto
E J
EJ
Instituto de Química y Fisicoquímica Biológicas (UBA-CONICET), Buenos Aires, Argentina.
Caamaño
C A
CA
Fernández
H N
HN
Dellacha
J M
JM
D000818:Animals; D001667:Binding, Competitive; D003432:Cross-Linking Reagents; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D013006:Growth Hormone / Q000378:metabolism / Q000031:analogs & derivatives; D007425:Intracellular Membranes / Q000032:analysis; D008099:Liver / Q000032:analysis* / Q000648:ultrastructure; D008862:Microsomes, Liver / Q000032:analysis* / Q000378:metabolism; D008970:Molecular Weight; D010782:Photolysis; D051381:Rats; D011919:Rats, Inbred Strains; D011981:Receptors, Prolactin / Q000032:analysis* / Q000378:metabolism; D011986:Receptors, Somatotropin / Q000032:analysis* / Q000378:metabolism; D012966:Sodium Cyanide / Q000494:pharmacology; D012995:Solubility
D003432:Cross-Linking Reagents; D011981:Receptors, Prolactin; D011986:Receptors, Somatotropin; C061509:somatotropin, di-(4-azidophenacyl) (182,189)-; D013006:Growth Hormone; D012966:Sodium Cyanide