title: Identification and characterization of binding proteins for inhibin and activin in human serum and follicular fluids.
abstract: Inhibins and activins are produced by a variety of tissues and may have important endocrine and paracrine roles in development, reproduction, and hematopoiesis. However, little is known regarding the physical properties or concentrations of inhibin and activin in biological fluids. Binding proteins for inhibin or activin in serum or at production or target sites may have important implications for restricting the bioactivity of these hormones and may alter the immunoreactivity of these molecules in biological fluids. The objective of this study was to identify inhibin- and activin-binding proteins in human serum (HS) and follicular fluid (hFF) and determine the ability of these proteins to alter biological or immunological activity. In HS, [125I]activin and inhibin bound to a protein identified as alpha 2-macroglobulin (alpha 2M) using three criteria: 1) [125I]inhibin and activin bind purified alpha 2M, but not several other serum proteins tested; 2) complexes formed by [125I]inhibin and activin in HS and in the presence of purified alpha 2M elute with similar retention times on HPLC; and 3) preadsorption of HS with alpha 2M antiserum inhibits inhibin and activin binding to this protein while antiserum directed against follistatin or other serum proteins had no effect. A small amount of a lower mol wt [125I]activin-follistatin complex was also found in HS. This complex eluted with a retention time similar to that of activin bound to purified porcine follistatin. Binding of inhibin to follistatin could not be detected in HS. In contrast, follistatin was the major binding protein of both activin and inhibin in hFF. Concentrations up to 100 micrograms/ml purified alpha 2M had no effect on the bioactivity or immunoreactivity of either inhibin or activin. In contrast, follistatin inhibited both activin-stimulated pituitary FSH release and K562 hemoglobin production as well as antiserum binding in a specific activin-A immunoassay. Follistatin did not interfere with inhibin immunodetection. These data indicate that two inhibin- and activin-binding proteins are present in different relative amounts in HS and hFF, alpha 2M, the primary binding protein in HS, did not alter inhibin or activin bio- or immunoactivity under the conditions of these experiments, while follistatin, the major binding protein in hFF, may mask activin's bio- and immunoactivities.
authors.affiliation: Genetech, Inc., South San Francisco, California 94080.
authors.forename: L A
authors.initials: LA
authors.lastname: Krummen
authors.forename: T K
authors.initials: TK
authors.lastname: Woodruff
authors.forename: G
authors.initials: G
authors.lastname: DeGuzman
authors.forename: E T
authors.initials: ET
authors.lastname: Cox
authors.forename: D L
authors.initials: DL
authors.lastname: Baly
authors.forename: E
authors.initials: E
authors.lastname: Mann
authors.forename: S
authors.initials: S
authors.lastname: Garg
authors.forename: W L
authors.initials: WL
authors.lastname: Wong
authors.forename: P
authors.initials: P
authors.lastname: Cossum
authors.forename: J P
authors.initials: JP
authors.lastname: Mather
mesh_terms: D028341:Activins; D000818:Animals; D002352:Carrier Proteins / Q000032:analysis* / Q000097:blood; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D005260:Female; D005640:Follicle Stimulating Hormone / Q000378:metabolism; D015571:Follicular Fluid / Q000737:chemistry*; D038681:Follistatin; D006023:Glycoproteins / Q000032:analysis / Q000378:metabolism / Q000494:pharmacology; D006454:Hemoglobins / Q000096:biosynthesis; D006801:Humans; D007163:Immunosorbent Techniques; D007265:Inhibins / Q000378:metabolism* / Q000494:pharmacology; D010902:Pituitary Gland / Q000187:drug effects / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D000511:alpha-Macroglobulins / Q000032:analysis / Q000378:metabolism / Q000494:pharmacology
chemical_list: D002352:Carrier Proteins; D038681:Follistatin; D006023:Glycoproteins; D006454:Hemoglobins; D000511:alpha-Macroglobulins; D028341:Activins; D007265:Inhibins; D005640:Follicle Stimulating Hormone