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

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.

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