Lactogen receptors in rat Leydig cells: analysis of their structure with bifunctional cross-linking reagents.
[125I]Iodohuman GH was found to bind to receptors with specificity for lactogenic hormones in a Triton X-100 extract from Leydig cell membranes displaying an affinity constant of 3.8 X 10(9) M-1 and a binding capacity of 167 fmol/mg protein. Cross-linking of solubilized [125I]iodohuman GH-receptor complexes with disuccinimidyl suberate followed by analysis by sodium dodecyl sulfate-gel electrophoresis in the presence of beta-mercaptoethanol and autoradiography resulted in the appearance of bands with apparent mol wt of 113,000, 103,000, 59,000, and 53,000. The appearance of these bands was prevented by incubation in the presence of lactogenic hormones. By using a two-dimensional electrophoresis technique (first dimension under nonreducing conditions; second dimension under reducing conditions), it was demonstrated that a fraction of the mol wt 59,000 species can be released from the mol wt 103,000 species upon cleavage of disulfide bonds. These results suggest the existence of lactogen receptor species with approximate mol wt of 91,000, 81,000, 37,000, and 31,000 in Triton X-100 extracts from Leydig cell membranes if the contribution of the free hormone (mol wt, 22,000) is subtracted. A fraction of the mol wt 37,000 subunits appears to be contained within the 81,000 species linked through disulfide bonds.
Bonifacino
J S
JS
Dufau
M L
ML
D000818:Animals; D055598:Chemical Phenomena; D002621:Chemistry; D003432:Cross-Linking Reagents; D004586:Electrophoresis; D013006:Growth Hormone / Q000378:metabolism; D006801:Humans; D007457:Iodine Radioisotopes; D007985:Leydig Cells / Q000378:metabolism*; D008297:Male; D051381:Rats; D011956:Receptors, Cell Surface / Q000378:metabolism*; D011981:Receptors, Prolactin; D012995:Solubility
D003432:Cross-Linking Reagents; D007457:Iodine Radioisotopes; D011956:Receptors, Cell Surface; D011981:Receptors, Prolactin; D013006:Growth Hormone