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Updated: Aug 15, 2026
Identifying Protein-protein Interaction Sites Using Peptide Arrays
Published on: November 18, 2014
Surface plasmon resonance imaging-based protein arrays for high-throughput screening of protein-protein interaction
Sun Ok Jung1, Hyeon-Su Ro, Byung Hoon Kho
1BioNanotechnology Research Center, Korea Research Institute of Bioscience and Biotechnology, Yuseong, Daejeon.
Abstract:
The E7 protein produced by high-risk human papillomavirus (HPV) induces a degradation of the retinoblastoma tumor suppressor RB through direct interaction, which suggests that an inhibitor for the interaction can be a potential anticancer drug. A surface plasmon resonance (SPR) imaging-based protein array chip was developed for the high-throughput screening of inhibitor molecules targeting RB-E7 interaction. The glutathione S-transferase-fused E7 protein (GST-E7) was first layered onto a glutathionylated gold chip surface that had been designed to specifically bind to GST-fused proteins. Subsequently, a microarrayer was used to spot the hexa-histidine-tagged RB proteins (His(6)-RB) onto the GST-E7-layered gold chip surface, and the resulting SPR image was analyzed. Upon increased His(6)-RB concentration in the spotting solution, the SPR signal intensity increased proportionally, indicating that His(6)-RB bound to GST-E7 in a concentration-dependent manner. The His(6)-RB/GST-E7 interaction was challenged by spotting the His(6)-RB solution in the presence of a RB binding peptide (PepC) derived from a motif on E7. The SPR imaging data showed that PepC inhibited the His(6)-RB/GST-E7 interaction in a concentration-dependent manner. Our results show that the SPR imaging-based protein array chip can be applied to screen small molecule inhibitors that target protein-protein interaction.
Insights
Researchers developed a novel protein array chip using surface plasmon resonance (SPR) imaging to screen for potential anticancer drugs that inhibit the human papillomavirus (HPV) E7 protein interaction with the retinoblastoma tumor suppressor (RB). This high-throughput method enables rapid identification of small molecules targeting protein-protein interactions.
Area of Science:
- Biochemistry
- Molecular Biology
- Oncology
Background:
- High-risk human papillomavirus (HPV) E7 protein targets the retinoblastoma tumor suppressor (RB), promoting cancer development.
- Inhibiting the RB-E7 protein-protein interaction is a promising strategy for anticancer drug development.
Purpose of the Study:
- To develop and validate a high-throughput screening method for identifying inhibitors of the RB-E7 interaction.
- To demonstrate the utility of a surface plasmon resonance (SPR) imaging-based protein array chip for drug discovery.
Main Methods:
- A gold chip was functionalized for immobilization of glutathione S-transferase-fused E7 protein (GST-E7).
- Hexa-histidine-tagged RB proteins (His(6)-RB) were spotted onto the GST-E7 layer using a microarrayer.
- SPR imaging was employed to analyze the binding kinetics and inhibition of the His(6)-RB/GST-E7 interaction.
Main Results:
- His(6)-RB demonstrated concentration-dependent binding to GST-E7 on the SPR chip.
- A peptide inhibitor (PepC) derived from E7 significantly inhibited the His(6)-RB/GST-E7 interaction in a concentration-dependent manner.
- The SPR imaging protein array chip successfully detected inhibition of the target protein-protein interaction.
Conclusions:
- The developed SPR imaging-based protein array chip is effective for high-throughput screening of small molecule inhibitors targeting protein-protein interactions.
- This platform holds significant potential for accelerating the discovery of novel anticancer therapeutics against HPV-driven cancers.
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