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Effect of receptor phosphorylation on the binding between IRS‐1 and IGF‐1R as revealed by surface plasmon resonance biosensor
Author(s) -
Huang Minghui,
Lai Wan Ping,
Wong Man Sau,
Yang Mengsu
Publication year - 2001
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(01)02780-6
Subject(s) - surface plasmon resonance , receptor , biosensor , phosphorylation , insulin receptor , irs1 , biophysics , extracellular , chemistry , intracellular , cell surface receptor , insulin receptor substrate , ligand (biochemistry) , biochemistry , insulin , biology , materials science , endocrinology , insulin resistance , nanoparticle , nanotechnology
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)×10 9 M −1 and (9.81±4.61)×10 8 M −1 , respectively.