Heparin-binding mechanism of the IGF2/IGF-binding protein 2 complex

Jacob Lund, Mads T. Søndergaard, Cheryl A Conover, Michael T. Overgaard

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

IGF1 and IGF2 are potent stimulators of diverse cellular activities such as differentiation and mitosis. Six IGF-binding proteins (IGFBP1-IGFBP6) are primary regulators of IGF half-life and receptor availability. Generally, the binding of IGFBPs inhibits IGF receptor activation. However, it has been shown that IGFBP2 in complex with IGF2 (IGF2/IGFBP2) stimulates osteoblast function in vitro and increases skeletal mass in vivo. IGF2 binding to IGFBP2 greatly increases the affinity for 2- or 3-carbon O-sulfated glycosaminoglycans (GAGs), e.g. heparin and heparan sulfate, which is hypothesized to preferentially and specifically target the IGF2/IGFBP2 complex to the bone matrix. In order to obtain a more detailed understanding of the interactions between the IGF2/IGFBP2 complex and GAGs, we investigated heparin-binding properties of IGFBP2 and the IGF2/IGFBP2 complex in a quantitative manner. For this study, we mutated key positively charged residues within the two heparin-binding domains (HBDs) in IGFBP2 and in one potential HBD in IGF2. Using heparin affinity chromatography, we demonstrate that the two IGFBP2 HBDs contribute differentially to GAG binding in free IGFBP2 and the IGF2/IGFBP2 protein complex. Moreover, we identify a significant contribution from the HBD in IGF2 to the increased IGF2/IGFBP2 heparin affinity. Using molecular modeling, we present a novel model for the IGF2/IGFBP2 interaction with heparin where all three proposed HBDs constitute a positively charged and surface-exposed area that would serve to promote the increased heparin affinity of the complex compared with free intact IGFBP2.

Original languageEnglish (US)
Pages (from-to)345-355
Number of pages11
JournalJournal of Molecular Endocrinology
Volume52
Issue number3
DOIs
StatePublished - Mar 6 2014

Fingerprint

Insulin-Like Growth Factor Binding Protein 2
Heparin
Insulin-Like Growth Factor Binding Proteins
A73025
Glycosaminoglycans
Bone Matrix
Heparitin Sulfate
Osteoblasts
Affinity Chromatography
Mitosis
Half-Life
Carbon

Keywords

  • Glycosaminoglycans
  • Heparin affinity
  • IGF2/IGFBP2 complex
  • Ternary binding model

ASJC Scopus subject areas

  • Endocrinology
  • Molecular Biology
  • Medicine(all)

Cite this

Heparin-binding mechanism of the IGF2/IGF-binding protein 2 complex. / Lund, Jacob; Søndergaard, Mads T.; Conover, Cheryl A; Overgaard, Michael T.

In: Journal of Molecular Endocrinology, Vol. 52, No. 3, 06.03.2014, p. 345-355.

Research output: Contribution to journalArticle

Lund, Jacob ; Søndergaard, Mads T. ; Conover, Cheryl A ; Overgaard, Michael T. / Heparin-binding mechanism of the IGF2/IGF-binding protein 2 complex. In: Journal of Molecular Endocrinology. 2014 ; Vol. 52, No. 3. pp. 345-355.
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