Receptor-binding activity of highly purified bovine luteinizing hormone and thyrotropin, and their subunits.
Highly purified preparations of bovine TSH (bTSH) and LH (bLH) and their subunits have been obtained by affinity chromatography using immobilized antibodies directed against counterpart subunits. The purified preparations were assessed for biological activity in radioligand-receptor assays for TSH and LH. After affinity purification against bLH beta, a TSH preparation whose initial potency in the LH assay had been 0.15% that of LH, failed to compete with [125I]LH in amounts up to 100 microgram. Thus, it appears that bTSH does not bind to LH receptors in the rat testis and that interaction of less purified TSH with gonadotropin receptors is attributable to LH contamination. In contrast, LH, whose initial potency in the TSH receptor assay was 0.6% that of TSH, retained a potency of 0.004% of TSH (equivalent to 3.6 mU/mg) after immunoadsorption by anti-bTSH beta. The retention of TSH receptor-binding activity by affinity-purified LH indicates that the LH molecule (like hCG) has a low intrinsic thyroid-stimulating activity. Affinity-purified LH subunits have little or no demonstrable affinity for the LH receptor in vitro. Affinity-purified TSH subunits and affinity-purified LH, however, exhibit very weak receptor-binding activity in the TSH radioligand receptor assay. An evaluation of the capacity of the immunoadsorbents to remove TSH from artificial mixtures suggests that the residual binding does not result entirely from contamination, and therefore, that alpha-subunits as well as LH have some intrinsic TSH-binding activity.
D000818:Animals; D002417:Cattle; D002846:Chromatography, Affinity; D007457:Iodine Radioisotopes; D007700:Kinetics; D007986:Luteinizing Hormone / Q000302:isolation & purification / Q000378:metabolism*; D046911:Macromolecular Substances; D011485:Protein Binding; D011869:Radioligand Assay; D011956:Receptors, Cell Surface / Q000378:metabolism*; D013552:Swine; D013961:Thyroid Gland / Q000378:metabolism; D013972:Thyrotropin / Q000302:isolation & purification / Q000378:metabolism*
D007457:Iodine Radioisotopes; D046911:Macromolecular Substances; D011956:Receptors, Cell Surface; D007986:Luteinizing Hormone; D013972:Thyrotropin