Extracellular matrix contains insulin-like growth factor binding protein-5: potentiation of the effects of IGF-I.
Insulin-like growth factor binding proteins (IGFBPs) have been shown to serve as carrier proteins for the insulin-like growth factors (IGFs) and to modulate their biologic effects. Since extracellular matrix (ECM) has been shown to be a reservoir for IGF-I and IGF-II, we examined the ECM of cultured human fetal fibroblasts and found that IGFBP-5 was incorporated intact into ECM, while mostly inert proteolytic fragments were found in the medium. In contrast, two other forms of IGFBP that are secreted by these cells were either present in ECM in minimal amounts (IGFBP-3) or not detected (IGFBP-4). Likewise, when purified IGFBPs were incubated with ECM, IGFBP-5 bound preferentially. IGFBP-5 was found to bind to types III and IV collagen, laminin, and fibronectin. Increasing salt concentrations inhibited the binding of IGFBP-5 to ECM and accelerated the release of IGFBP-5 from ECM, suggesting an ionic basis for this interaction. ECM-associated IGFBP-5 had a sevenfold decrease in affinity for IGF-I compared to IGFBP-5 in solution. Furthermore, when IGFBP-5 was present in cell culture substrata, it potentiated the growth stimulatory effects of IGF-I on fibroblasts. When IGFBP-5 was present only in the medium, it was degraded to a 22-kD fragment and had no effect on IGF-I-stimulated growth. We conclude that IGFBP-5 is present in fibroblast ECM, where it is protected from degradation and can potentiate the biologic actions of IGF-I. These findings provide a molecular explanation for the association of the IGF's with the extracellular matrix, and suggest that the binding of the IGF's to matrix, via IGFBP-5, may be important in mediating the cellular growth response to these growth factors.
Jones
J I
JI
Department of Medicine, University of North Carolina School of Medicine, Chapel Hill 27599-7170.
Gockerman
A
A
Busby
W H
WH
Camacho-Hubner
C
C
Clemmons
D R
DR
D001667:Binding, Competitive; D002352:Carrier Proteins / Q000378:metabolism / Q000494:pharmacology*; D002478:Cells, Cultured / Q000187:drug effects; D003094:Collagen / Q000378:metabolism; D004357:Drug Synergism; D005109:Extracellular Matrix / Q000737:chemistry*; D005347:Fibroblasts / Q000187:drug effects; D005353:Fibronectins / Q000378:metabolism; D006801:Humans; D018975:Insulin-Like Growth Factor Binding Protein 5; D007334:Insulin-Like Growth Factor I / Q000494:pharmacology*; D007477:Ions; D007797:Laminin / Q000378:metabolism; D012867:Skin / Q000166:cytology; D013002:Somatomedins / Q000378:metabolism / Q000494:pharmacology*
D002352:Carrier Proteins; D005353:Fibronectins; D018975:Insulin-Like Growth Factor Binding Protein 5; D007477:Ions; D007797:Laminin; D013002:Somatomedins; D007334:Insulin-Like Growth Factor I; D003094:Collagen