Identification of a luteotrophic protein in bovine allantoic fluid

Allantoic fluids (n = 65) of Day 24-37 bovine conceptuses were collected and assayed for luteotrophic activity in vitro with dispersed bovine luteal cells. Significant luteotrophic activity was found in 41% of the samples, with the highest percentage occurring between Days 25 and 28. The activity is ammonium sulphate-precipitable, heat-labile and inactivated by trypsin and chymotrypsin. Gel filtration column chromatography identified one peak of luteotrophic activity with a molecular weight of 68,000. Concanavalin A bound the luteotrophic activity, thus allowing rapid and substantial purification from a major protein of Mr 68,000 which was concanavalin A non-reactive. The results of one- and two-dimensional SDS-PAGE of concanavalin A-reactive fractions containing activity suggest that the luteotrophic activity present in allantoic fluid is associated with a glycoprotein of Mr 68,000 present in very low concentrations. The active factor does not appear to be alpha-fetoprotein. This protein may be an important conceptus-derived luteotrophin that stimulates progesterone production by the corpus luteum of cows during pregnancy.