Inhibin forms in human plasma.
In order to identify the molecular weight forms of bioactive and immunoactive inhibin in human plasma, plasma/serum was sequentially fractionated by immunoaffinity chromatography (using immobilised inhibin alpha subunit antiserum), reversed phase HPLC and preparative SDS-PAGE. The electroeluted gel fractions were assayed for inhibin in vitro bioactivity and immunoactivity, the latter by RIA. Initial experiments examined human follicular fluid as an inhibin-rich source. Bioactive and immunoactive fractions of 30, 35, 53, 65 and approximately 120 kDa were identified in addition to bio-inactive, immunoactive fractions of 26 kDa and 32 kDa. These molecular weights correspond to those of known inhibin forms and are attributed to differing degrees of glycosylation of the inhibin alpha subunit and variable processing of the alpha and beta inhibin subunits. Fractionation of male plasma pools revealed the presence of higher molecular weight immunoactive forms (55-120 kDa) as well as 28-31 kDa forms although the molecular weight distribution of activity between pools varied. To assess if the molecular weight pattern was modified by storage and/or subsequent fractionation, protease inhibitors were added initially to plasma and fractionated as above. The molecular weight distribution of immunoactivity was largely unaffected by the treatment, indicating that minimal processing had occurred. Postmenopausal serum itself showed low to undetectable activity. The addition of recombinant human 31 kDa inhibin to postmenopausal serum resulted in a molecular weight profile of inhibin immunoactivity consistent with the presence of 31 kDa inhibin. Fractionation of a serum pool from women undergoing gonadotrophin stimulation, in which inhibin levels were elevated, showed a range of bioactive and immunoactive inhibin forms over the 30-120 kDa range.(ABSTRACT TRUNCATED AT 250 WORDS)
Robertson
D M
DM
Prince Henry's Institute of Medical Research, Monash Medical Centre, Clayton, Victoria, Australia.
Sullivan
J
J
Watson
M
M
Cahir
N
N
D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D007265:Inhibins / Q000097:blood* / Q000737:chemistry; D008297:Male; D008970:Molecular Weight; D011863:Radioimmunoassay
D007265:Inhibins