Identification of inhibin and inhibin-related proteins in human follicular fluid.
Inhibin-related proteins were identified in human follicular fluid following fractionation by gel permeation chromatography under neutral and acidic conditions, reversed-phase high performance liquid chromatography (HPLC) and preparative polyacrylamide gel electrophoresis. A number of molecular mass forms of inhibin (30-36 and 59-66 kDa) based on their in vitro biological and immunological activities were identified, of which 59-66 kDa inhibin was the predominant form. Bioactive fractions devoid of inhibin immunoactivity were also identified with molecular masses of 46 and 55 kDa. Based on their retention positions on reversed-phase HPLC and their lack of inhibin immunoactivity, these proteins are likely to be follicle stimulating hormone (FSH) suppressing proteins/follistatins previously identified in bovine and porcine follicular fluids. In addition, an immunoactive inhibin fraction devoid of bioactivity was identified in large amounts in the 50-70 kDa region of the gel permeation chromatogram at neutral pH. This material, based on previous findings with the fractionation of bovine follicular fluid, is likely to be the alpha-inhibin subunit precursor protein. No FSH stimulating activity (activin) was identified in any of the chromatograms, suggesting that the levels of activin in human follicular fluid are low. In conclusion, inhibins of molecular mass 30, 36, 59 and 66 kDa have been identified in human follicular fluid. Proteins with inhibin bioactivity devoid of immunoactivity and vice versa have also been detected and these proteins are probably FSH suppressing protein and an alpha-inhibin subunit precursor protein. Activin could not be identified.
Robertson
D M
DM
Department of Anatomy, Monash University, Clayton, Vic., Australia.
Foulds
L M
LM
de Vos
F
F
Leversha
L
L
de Kretser
D M
DM
D005591:Chemical Fractionation; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D015571:Follicular Fluid / Q000737:chemistry*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007265:Inhibins / Q000032:analysis*
D007265:Inhibins