Beattie James, Phillips Kirsten, Shand John H, Szymanowska Malgorzata, Flint David J, Allan Gordon J
Strathclyde Institute of Pharmacy & Biomedical Science, Royal College Building, University of Strathclyde, Glasgow, UK.
Mol Cell Biochem. 2008 Jan;307(1-2):221-36. doi: 10.1007/s11010-007-9601-8. Epub 2007 Sep 25.
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.
本综述描述了基于表面等离子体共振（SPR）的生物传感器对胰岛素样生长因子（IGF）分子轴中分子相互作用的综合分析。在本研究中，我们重点关注多肽生长因子IGF-I和IGF-II与六种可溶性IGF结合蛋白（IGFBP 1-6）之间的相互作用，这些蛋白天然存在于各种生物体液中。我们描述了准确测定这些相互作用动力学速率常数所需的条件，并强调了在此类研究早期可能遇到的实验和理论陷阱。我们重点研究IGFBP-5，并描述了一项定点诱变研究，该研究考察了蛋白质中各种残基对与IGF-I和-II高亲和力相互作用的贡献。我们分析了IGFBP-5（和IGFBP-3）与肝素及其他生物分子的相互作用，并描述了旨在监测该分子轴中多蛋白复合物形成的实验。
