Isolation and partial characterization of a Mr 32,000 protein with inhibin activity from porcine follicular fluid

Abstract

A Mr 32,000 protein with inhibin activity was isolated from porcine follicular fluid by heparin-Sepharose affinity chromatography, gel filtration on Sephacryl S-200, and four reversed-phase HPLC steps. The isolated molecule is composed of two chains having molecular weights of 18,000 and 14,000, respectively, and bound together by disulfide bonds. Amino acid sequence analysis revealed the 10 NH2-terminal residues of the Mr 18,000 chain to be Ser-Thr-Ala-Pro-Leu-Pro-Trp-Pro-Trp-Ser- and those of the Mr 14,000 chain to be Gly-Leu-Glu-Xaa-Asp-Gly-Arg-Thr-Asn-Leu-. This Mr 32,000 protein specifically inhibits the basal secretion of FSH, but not that of LH, in the rat anterior pituitary monolayer culture system, with a half-maximal effective dose of 450 pg/ml.