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Surface plasmon resonance imaging‐based protein arrays for high‐throughput screening of protein‐protein interaction inhibitors
Author(s) -
Jung Sun Ok,
Ro HyeonSu,
Kho Byung Hoon,
Shin YongBeom,
Kim Min Gon,
Chung Bong Hyun
Publication year - 2005
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200500001
Subject(s) - surface plasmon resonance , protein–protein interaction , chemistry , protein microarray , small molecule , retinoblastoma protein , target protein , peptide , biophysics , biochemistry , nanotechnology , biology , materials science , nanoparticle , dna microarray , gene expression , cell cycle , cell , gene
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.