Molecular interactions in the insulin-like growth factor (IGF) axis: a surface plasmon resonance (SPR) based biosensor study.
This review describes a comprehensive analysis of a surface plasmon resonance (SPR)-based biosensor study of molecular interactions in the insulin-like growth factor (IGF) molecular axis. In this study, we focus on the interaction between the polypeptide growth factors IGF-I and IGF-II with six soluble IGF binding proteins (IGFBP 1-6), which occur naturally in various biological fluids. We have describe the conditions required for the accurate determination of kinetic rate constants for these interactions and highlight the experimental and theoretical pitfalls, which may be encountered in the early stages of such a study. We focus on IGFBP-5 and describe a site-directed mutagenesis study, which examines the contribution of various residues in the protein to high affinity interaction with IGF-I and -II. We analyse the interaction of IGFBP-5 (and IGFBP-3) with heparin and other biomolecules and describe experiments, which were designed to monitor multi-protein complex formation in this molecular axis.
Beattie
James
J
Strathclyde Institute of Pharmacy & Biomedical Science, Royal College Building, University of Strathclyde, Glasgow, UK. james.beattie@strath.ac.uk
Phillips
Kirsten
K
Shand
John H
JH
Szymanowska
Malgorzata
M
Flint
David J
DJ
Allan
Gordon J
GJ
D000595:Amino Acid Sequence; D000818:Animals; D015374:Biosensing Techniques / Q000379:methods*; D006493:Heparin / Q000032:analysis / Q000378:metabolism; D006801:Humans; D018975:Insulin-Like Growth Factor Binding Protein 5 / Q000235:genetics; D018969:Insulin-Like Growth Factor Binding Proteins / Q000378:metabolism*; D008024:Ligands; D008969:Molecular Sequence Data; D016296:Mutagenesis / Q000502:physiology; D011485:Protein Binding; D013002:Somatomedins / Q000032:analysis / Q000378:metabolism* / Q000502:physiology*; D020349:Surface Plasmon Resonance / Q000379:methods*
D018975:Insulin-Like Growth Factor Binding Protein 5; D018969:Insulin-Like Growth Factor Binding Proteins; D008024:Ligands; D013002:Somatomedins; D006493:Heparin