PURIFICATION OF GONADOTROPIN SURGE-INHIBITING FACTOR FROM SERTOLI CELL-ENRICHED CULTURE-MEDIUM

Gonadotropin surge-inhibiting factor (GnSIF) is a novel biological factor in follicular fluid that inhibits the pre-ovulatory LH and FSH surges. Recent studies suggest that the biological activity of GnSIF can be attributed to inhibin. Preliminary studies showed that the Sertoli cell-conditioned medium contains GnSIF-like biological activity. Using 32 liters of primary Sertoli cell-enriched culture medium and an in vitro pituitary bioassay, GnSIF was isolated. GnSIF is a single monomeric polypeptide (Mr of 37,000) with a partial N-terminal amino acid sequence of NH2-SDXXPQL which is distinct from any existing protein sequences. The identity of the inhibin that was separated from GnSIF was confirmed by direct protein sequencing. The purified GnSIF inhibited GnRH-stimulated LH- and basal FSH-release from pituitary cells cultured in vitro dose-dependently with a half maximal effective dose of about 1.4 ng/ml and 3.5 ng/ml, respectively. The half maximal effective dose of inhibin to suppress the pituitary GnRH-stimulated LH- and basal FSH-release was 2.5 ng/ml and 0.8 ng/ml, respectively. This study demonstrates that GnSIF is a unique protein that shares some biological activities with inhibin. (C) 1994 Academic Press, Inc.