Biological activity of partially purified low molecular weight luteinizing hormone binding inhibitor in porcine follicular fluids.
We performed partial purification of low molecular weight luteinizing hormone binding inhibitor from porcine follicular fluids and examined its biological activities. Following ultrafiltration, gel filtration and anion exchange of the pooled porcine follicular fluids, low molecular weight fractions (500-10,000 MW) inhibited [125I]hLH binding to porcine granulosa cells in a dose-dependent manner. The binding inhibition kinetics study revealed that the luteinizing hormone binding inhibitor may indicate a non-competitive inhibition with [125I]hLH binding. In vitro bioassay using adult mouse testicular interstitial cells revealed that the partially purified luteinizing hormone binding inhibitor reduced ovine LH-stimulated testosterone and cAMP production in a dose-dependent manner, whereas the luteinizing hormone binding inhibitor did not affect basal production of testosterone and cAMP. The inhibitory activity was heat stable and did not disappear with activated charcoal adsorption. The results of the present study suggest that the luteinizing hormone binding inhibitor may play an important role as an ovarian non-steroidal regulator modulating the receptor binding of LH and LH-mediated steroidogenesis.
Department of Obstetrics and Gynecology, Wakayama Medical College, Japan.
N
N
Kokawa
M
M
Yamoto
K
K
Furukawa
R
R
Nakano
D000818:Animals; D001681:Biological Assay; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D000242:Cyclic AMP / Q000096:biosynthesis; D004305:Dose-Response Relationship, Drug; D005260:Female; D015571:Follicular Fluid / Q000737:chemistry*; D036341:Intercellular Signaling Peptides and Proteins; D007986:Luteinizing Hormone / Q000494:pharmacology; D008297:Male; D010455:Peptides / Q000302:isolation & purification / Q000502:physiology*; D011956:Receptors, Cell Surface / Q000037:antagonists & inhibitors* / Q000302:isolation & purification; D011974:Receptors, LH / Q000378:metabolism; D013552:Swine; D013737:Testis / Q000378:metabolism; D013739:Testosterone / Q000096:biosynthesis
D036341:Intercellular Signaling Peptides and Proteins; C011898:LH receptor binding inhibitor; D010455:Peptides; D011956:Receptors, Cell Surface; D011974:Receptors, LH; D013739:Testosterone; D007986:Luteinizing Hormone; D000242:Cyclic AMP