Isolation and characterization of a gonadotropin receptor binding inhibitor from porcine follicular fluid.
The presence of a gonadotropin receptor binding inhibitor in pooled porcine follicular fluid has been demonstrated. Porcine follicular fluid fractionation on DE-32 at near neutral pH, followed by a cation exchange chromatography on SPC-50 and Cibacron blue affinity chromatography, yielded a partially purified gonadotropin receptor binding inhibitor (GI-4). The partially purified GI binding inhibitor inhibited the binding of both 125I labelled hFSH and hCG to rat ovarian receptor preparation. SDS electrophoresis of radioiodinated partially purified GI followed by autoradiography made it possible to identify the binding component as a protein of molecular weight of 80,000. Subjecting 125I labelled GI-4 to chromatography on Sephadex G-100 helped obtain a homogeneous material, GI-5. The 125I labelled GI-5 exhibited in its binding to ovarian membrane preparations characteristics typical of a ligand-receptor interaction such as saturability, sensitivity to reaction conditions as time, ligand and receptor concentrations and finally displaceability by unlabelled inhibitor as well as FSH and hCG in a dose dependent manner. This material could bind ovarian receptors for both FSH and LH, its binding being inhibited by added FSH or hCG in a dose dependent manner.
Bhat
P J
PJ
Department of Biochemistry, Indian Institute of Science, Bangalore.
Moudgal
N R
NR
D000818:Animals; D001667:Binding, Competitive; D001685:Biological Factors / Q000302:isolation & purification / Q000378:metabolism*; D005260:Female; D008970:Molecular Weight; D006080:Ovarian Follicle / Q000378:metabolism*; D011962:Receptors, FSH / Q000378:metabolism; D011967:Receptors, Gonadotropin / Q000378:metabolism*; D011974:Receptors, LH / Q000378:metabolism; D013552:Swine
D001685:Biological Factors; D011962:Receptors, FSH; D011967:Receptors, Gonadotropin; D011974:Receptors, LH