Previous studies demonstrated that IGF-II binds directly to vitronectin (VN) while IGF-I binds poorly. However, binding of VN to integrins has been demonstrated to be essential for a range of IGF-I-stimulated biological effects including IGF binding protein-5 (IGFBP-5) production, IGF type-1 receptor autophosphorylation and cell migration. Thus, this study examined the hypothesis that a link between IGF-I and VN must occur and may be mediated through IGFBPs. Studies using competitive binding assays with VN and [125I]-labelled IGFs in the absence and presence of IGFBPs revealed IGFBP-4, IGFBP-5 and non-glycoyslated IGFBP-3 significantly enhance binding of IGF-I to VN, while IGFBP-2 and glycosylated IGFBP-3 had a smaller effect. Furthermore, binding studies with analogues indicate that glycosylation status of IGFBP-3 and the heparin-binding domains of IGFBP-3 and IGFBP-5 are important in this interaction. The functional significance of IGFs binding to VN on cell migration in MCF-7 breast carcinoma cells was examined and cell migration was found to be enhanced when VN was pre-bound to IGF-I in the presence of IGFBP-3, -4 and -5. The effect required IGF:IGFBP:VN complex formation; this was demonstrated by use of a non-IGFBP-binding analogue, des(1- 3)IGF-I. Additionally, higher doses of IGFs in the presence of VN also could stimulate cell migration. Together, these data indicated the importance of IGFBPs in modulating IGF-I binding to VN and that this binding has functional consequences in cells. Future directions for this work include investigations into the mechanisms underlying formation of the trimeric complex and the associated signalling pathways involved.
| Identifer | oai:union.ndltd.org:ADTP/265057 |
| Date | January 2005 |
| Creators | Kricker, Jennifer Ann |
| Publisher | Queensland University of Technology |
| Source Sets | Australiasian Digital Theses Program |
| Detected Language | English |
| Rights | Copyright Jennifer Ann Kricker |
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