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Title 

Surface plasmon resonance imaging-based protein arrays for high-throughput screening of protein-protein interaction inhibitors

Authors 

Sun Ok JungH S RoByung Hoon KhoYong Beom ShinMin-Gon KimBong Hyun Chung

Publisher 

Wiley-Blackwell

Issue Date 

2005

Citation 

Proteomics, vol. 5, no. 17, pp. 4427-4431

Keywords 

high-throughput screeningprotein arraysprotein-protein interactionsurface plasmon resonance (SPR) imaginghybrid proteinprotein E7protein inhibitorretinoblastoma proteinhigh throughput screeningimaging

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 His6-RB concentration in the spotting solution, the SPR signal intensity increased proportionally, indicating that His6-RB bound to GST-E7 in a concentration-dependent manner. The His6-RB/GST-E7 interaction was challenged by spotting the His6-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 His6-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.

ISSN 

1615-9853

Link 

http://dx.doi.org/10.1002/pmic.200500001

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2017-04-19


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