Binding of Zona Binding Inhibitory Factor-1 (ZIF-1) from Human Follicular Fluid on Spermatozoa

Glycodelin-S in Human Seminal Plasma Reduces Cholesterol Efflux andInhibits Capacitation ofSpermatozoa

“…Radioactively labeled GdS was prepared by mixing 50 g of GdS in 0.02 ml of 0.05 M PBS (pH 7.4) with 2 mCi of sodium 125 I (20 l, Amersham Biosciences) and freshly prepared chloramine T (100 g in 0.02 ml of 0.05 M PBS, pH 7.4) as described (12). The first radioactive peak containing iodinated glycodelin was collected.…”

“…Deglycosylated GdS was prepared using the N-glycosidase F deglycosylation kit (Bio-Rad) as described (12). The deglycosylated protein was obtained after three successive protein precipitations, redissolved in 20 l of PBS, and further purified by gel filtration chromatography in a SMART system (Amersham Biosciences).…”

“…The processed spermatozoa were then resuspended in Earle's balanced salt solution supplemented with 0.3% BSA, 0.3 mM sodium pyruvate, 0.16 mM penicillin-G, 0.05 mM streptomycin sulfate, and 14 mM sodium bicarbonate (EBSS/0.3%BSA) to a concentration of 2 ϫ 10 6 spermatozoa/ml. Purification of Glycodelin Isoforms-Glycodelin-A (GdA), -S (GdS), and -F (GdF) were purified from amniotic fluid, seminal plasma, and follicular fluid respectively, using a monoclonal anti-glycodelin antibody (clone F43-7F9) Sepharose column as described (12). The bound glycodelin was eluted with 0.1% trifluoroacetic acid and dialyzed against 100 mM sodium phosphate buffer (pH 7.2).…”

“…The bound glycodelin was eluted with 0.1% trifluoroacetic acid and dialyzed against 100 mM sodium phosphate buffer (pH 7.2). GdS was further purified by anion exchange chromatography as described (12). GdF was purified from human follicular fluid collected during oocyte retrieval from women undergoing assisted reproduction treatment in Queen Mary Hospital, Hong Kong.…”

“…GdF also inhibits spermatozoa-ZP binding (8). GdF, but not the other glycodelin isoforms, suppresses progesterone-induced acrosome reaction (8,12), suggesting that it may protect spermatozoa from premature acrosome reaction before binding to the ZP. Glycodelin-S does not adversely affect spermatozoa-zona binding (6), and its action on spermatozoa is not clear.…”