Human follicular fluid contains a follicle-stimulating hormone (FSH) receptor binding inhibitor which has FSH agonist activity, is immunologically similar to FSH, but can be distinguished from FSH.
We have previously reported the presence in human follicular fluid (hFF) of a high (greater than 5000) mol wt FSH receptor binding inhibitor (FSH-BI). This hFF FSH-BI was further purified by removal of material insoluble in acidified acetone (pH 4.1) but soluble in diethyl ether (pH 10.5), followed by molecular sieving through Sephacryl S-100. FSH-BI activity eluted from S-100 with an elution volume similar to that of hFSH, but could be distinguished from hFSH on the basis of a differential sensitivity to acid inactivation. Human FSH was inactivated in acetone at pH 4.1 (1 h, 25 C), whereas hFF FSH-BI retained activity under these conditions. Human FF FSH-BI also demonstrated FSH-like agonist activity, defined as the ability to stimulate basal levels of estradiol synthesis in cultured rat Sertoli cells. Human FSH-BI strongly cross-reacted to a commercially available monoclonal antibody used to measure serum hFSH. Indeed, recovery of FSH immunologic activity was significantly greater (134-fold on a mass basis) after partial purification, indicating that antibody recognition sites were apparently masked in unfractionated hFF. In summary, large mol wt hFSH-BI has been partially purified from hFF and found to be similar in size to pituitary hFSH and to have FSH-like agonist activity in vitro. Although distinguishable from pituitary hFSH on the basis of stability to acid, hFSH-BI appears immunologically related to pituitary hFSH so that measurements of hFSH levels in hFF using immunologic techniques should be interpreted with caution.
Lee
D W
DW
Department of Biochemistry, Albany Medical College, New York 12208.
Butler
W J
WJ
Horvath
P M
PM
Shelden
R M
RM
Reichert
L E
LE
D000096:Acetone / Q000494:pharmacology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002352:Carrier Proteins; D002478:Cells, Cultured; D002850:Chromatography, Gel; D003429:Cross Reactions / Q000276:immunology; D004958:Estradiol / Q000096:biosynthesis; D005260:Female; D005640:Follicle Stimulating Hormone / Q000037:antagonists & inhibitors* / Q000276:immunology / Q000502:physiology; D015571:Follicular Fluid / Q000737:chemistry* / Q000502:physiology; D006020:Glycopeptides / Q000032:analysis* / Q000276:immunology / Q000502:physiology; D006801:Humans; D008297:Male; D011869:Radioligand Assay; D051381:Rats; D011919:Rats, Inbred Strains; D012708:Sertoli Cells / Q000187:drug effects / Q000378:metabolism
D000911:Antibodies, Monoclonal; D002352:Carrier Proteins; C484018:FSH-BI protein, human; D006020:Glycopeptides; C063151:follicle-stimulating hormone-suppressing protein, rat; D000096:Acetone; D004958:Estradiol; D005640:Follicle Stimulating Hormone