Activin-binding proteins in human serum and follicular fluid.
Binding proteins that transport and/or modify the biological action of peptide hormones and growth factors have been identified for an increasing number of endocrinologically important substances. Since these binding proteins can mask epitopes critical for recognition in immunoassays and can neutralize the bioactivity of their targets, elucidation of hormonal physiology can be intricately tied to analysis of binding protein structure and function. Therefore, we investigated whether circulating activin- and inhibin-binding proteins exist in human serum by incubating purified recombinant human 125I-activin with serum samples. After gel permeation chromatography, radioactive activin was identified in three peaks, a high molecular wt (mol wt) binding protein peak (230 kDa), a lower mol wt binding protein peak (60 kDa), and free activin (22.5 kDa). Bound activin was displaced from the lower mol wt binding protein with either activin or inhibin, but was not displaced from the high mol wt peak with a 10-fold greater concentration of activin. Since an activin-binding protein, follistatin, has been identified in ovarian and pituitary extracts, these same analytical techniques were applied to analysis of human follicular fluid as well. A large, 60 kDa binding protein peak eluting in a position similar to the lower mol wt peak in serum was observed, consistent with this protein being follistatin. These results demonstrate the presence of at least two activin-binding proteins, distinguishable by size, in human serum that may interfere with attempts to assay activin levels in circulation without prior extraction, and may also be involved in regulating the biological actions of activin.
Schneyer
A L
AL
Department of Medicine, Massachusetts General Hospital, Boston 02114.
O'Neil
D A
DA
Crowley
W F
WF
D028341:Activins; D000328:Adult; D002850:Chromatography, Gel; D005260:Female; D038681:Follistatin; D006023:Glycoproteins / Q000097:blood / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D007265:Inhibins / Q000378:metabolism*; D008297:Male; D008970:Molecular Weight; D006080:Ovarian Follicle / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism
D038681:Follistatin; D006023:Glycoproteins; D011994:Recombinant Proteins; D028341:Activins; D007265:Inhibins