Purification and characterization of ovarian LH/hCG and prolactin receptors.
We have purified the luteinizing hormone (LH)/human choriogonadotropin (hCG) receptor to homogeneity by sequential affinity column on wheat germ lectin-Sepharose and hCG-Sepharose. The method was designed to allow also the purification of lactogen receptor from the initial starting material. Comparable purification of lactogen receptor can be attained using Con A-Sepharose as initial step. The purified LH/hCG receptor was identified as a single protein of Mr = 75,000 on SDS gel electrophoresis. The lactogen receptor is composed of two dissimilar active subunits of Mr 88,000 and 40,000, the latter probably being an integral part of the larger form. Comparison of Mr's derived from SDS gels with those from fast performance liquid chromatography suggested that the native LH holoreceptor is present in a dimeric form, while the lactogen receptor seems to be composed of aggregates that could represent dimeric or trimeric forms of holoreceptor Mr 80,000. Cross-linking studies performed after binding of hCG (radiolabeled in the individual subunits) to the purified LH/hCG receptor indicated that the hCG alpha-subunit undergoes predominant interaction with the receptor molecule. The influence of the beta-subunit in this interaction seems to occur mainly through its association with the alpha-subunit, presumably by conferring specificity to the alpha-subunit for its interaction with the receptor. The alpha-subunit, which is identical within species, has an important role in the receptor binding interaction and biological activity of glycoprotein hormones.
D000818:Animals; D001665:Binding Sites; D006063:Chorionic Gonadotropin / Q000378:metabolism*; D002846:Chromatography, Affinity; D005260:Female; D007986:Luteinizing Hormone / Q000378:metabolism*; D008970:Molecular Weight; D010053:Ovary / Q000032:analysis; D011388:Prolactin / Q000378:metabolism*; D011485:Protein Binding; D051381:Rats; D011974:Receptors, LH / Q000302:isolation & purification* / Q000378:metabolism; D011981:Receptors, Prolactin / Q000302:isolation & purification* / Q000378:metabolism
D006063:Chorionic Gonadotropin; D011974:Receptors, LH; D011981:Receptors, Prolactin; D011388:Prolactin; D007986:Luteinizing Hormone