Effect of receptor phosphorylation on the binding between IRS‐1 and IGF‐1R as revealed by surface plasmon resonance biosensor

FEBS Letters

Huang, Minghui²  Lai, Wan Ping¹  Wong, Man Sau¹  Yang, Mengsu² 

[1] Open Laboratory of Asymmetric Synthesis, Department of Applied Biology and Chemical Technology, Hong Kong Polytechnic University, Kowloon, Hong Kong, PR China;Applied Research Center for Genomic Technology, and Department of Biology and Chemistry, City University of Hong Kong, 83 Tat Chee Avenue, Kowloon, Hong Kong, PR China

关键词: Insulin-like growth factor-1 receptor;    Insulin receptor substrate-1;    Tyrosine phosphorylation;    Surface plasmon resonance;    Biosensor;    IGF-1;    insulin-like growth factor-1;    IGF-1R;    insulin-like growth factor-1 receptor;    IRS-1;    insulin receptor substrate-1;    SPR;    surface plasmon resonance;   

DOI  :  10.1016/S0014-5793(01)02780-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

A receptor binding assay based on the surface plasmon resonance (SPR) biosensor technique was developed to study the interaction between insulin-like growth factor-1 receptor (IGF-1R) and its intracellular substrate protein insulin receptor substrate-1 (IRS-1). The sensor surface was modified with anti-IGF-1R (α-subunit) monoclonal antibodies for the capturing of the receptor-containing membrane fragments from cell lysates. The IGF-1R was successfully immobilized on the sensor surface with binding capability for its intracellular substrates. SPR measurements showed that the tyrosine phosphorylation of IGF-1R induced by its extracellular ligand insulin-like growth factor-1 caused the receptor to bind with IRS-1 10 times faster than the unactivated receptor. As a result, the affinity constants of IRS-1 to phosphorylated and unphosphorylated IGF-1R were (8.06±5.18)×10⁹ M⁻¹ and (9.81±4.61)×10⁸ M⁻¹, respectively.

【 授权许可】

Unknown