Abstract
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) × 109M-1and (9.81 ± 4.61) × 108M-1, respectively. on behalf of the Federation of European Biochemical Societies.
| Original language | English |
|---|---|
| Pages (from-to) | 31-36 |
| Number of pages | 6 |
| Journal | FEBS Letters |
| Volume | 505 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - 7 Sept 2001 |
Keywords
- Biosensor
- Insulin receptor substrate-1
- Insulin-like growth factor-1 receptor
- Surface plasmon resonance
- Tyrosine phosphorylation
ASJC Scopus subject areas
- Biochemistry
- Biophysics
- Molecular Biology