title: Biological activity of follistatin isoforms and follistatin-like-3 is dependent on differential cell surface binding and specificity for activin, myostatin, and bone morphogenetic proteins.
abstract: Follistatin (FST) and FST-like-3 (FSTL3) are activin-binding and neutralization proteins that also bind myostatin. Three FST isoforms have been described that differ in tissue distribution and cell-surface binding activity, suggesting that the FST isoforms and FSTL3 may have some nonoverlapping biological actions. We produced recombinant FST isoforms and FSTL3 and compared their biochemical and biological properties. Activin-binding affinities and kinetics were comparable between the isoforms and FSTL3, whereas cell-surface binding differed markedly (FST288 > FST303 > FST315 > FSTL3). Inhibition of endogenous activin bioactivity, whether the FST isoforms were administered endogenously or exogenously, correlated closely with surface binding activity, whereas neutralization of exogenous activin when FST and FSTL3 were also exogenous was consistent with their equivalent activin-binding affinities. This difference in activin inhibition was also evident in an in vitro bioassay because FST288 suppressed, whereas FST315 enhanced, activin-dependent TT cell proliferation. Moreover, when FSTL3, which does not associate with cell membranes, was expressed as a membrane-anchored protein, its endogenous activin inhibitory activity was dramatically increased. In competitive binding assays, myostatin was more potent than bone morphogenetic proteins (BMPs) 6 and 7, and BMPs 2 and 4 were inactive in binding to FST isoforms, whereas none of the BMPs tested competed with activin for binding to FSTL3. Neutralization of exogenous BMP or myostatin bioactivity correlated with the relative abilities of the isoforms to bind cell-surface proteoglycans. These results indicate that the differential biological actions among the FST isoforms and FSTL3 are primarily dependent on their relative cell-surface binding ability and ligand specificity.
authors.affiliation: Reproductive Endocrine Unit BHX-5, Massachusetts General Hospital, Boston, 02114, USA.
authors.forename: Yisrael
authors.initials: Y
authors.lastname: Sidis
authors.forename: Abir
authors.initials: A
authors.lastname: Mukherjee
authors.forename: Henry
authors.initials: H
authors.lastname: Keutmann
authors.forename: Anne
authors.initials: A
authors.lastname: Delbaere
authors.forename: Miyuki
authors.initials: M
authors.lastname: Sadatsuki
authors.forename: Alan
authors.initials: A
authors.lastname: Schneyer
mesh_terms: D028341:Activins / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D019485:Bone Morphogenetic Proteins / Q000378:metabolism*; D019556:COS Cells; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D049109:Cell Proliferation; D002522:Chlorocebus aethiops; D038681:Follistatin / Q000737:chemistry* / Q000378:metabolism; D038702:Follistatin-Related Proteins / Q000378:metabolism*; D006801:Humans; D055435:Myostatin; D011994:Recombinant Proteins / Q000737:chemistry; D016212:Transforming Growth Factor beta / Q000378:metabolism*
chemical_list: D019485:Bone Morphogenetic Proteins; D038681:Follistatin; D038702:Follistatin-Related Proteins; C525911:MSTN protein, human; D055435:Myostatin; D011994:Recombinant Proteins; D016212:Transforming Growth Factor beta; D028341:Activins