Versatile analysis of multiple macromolecular interactions by SPR imaging: application to p53 and DNA interaction.
The greatest challenge in the postgenomic era is the description of proteome interactions, such as protein-protein or protein-DNA interactions. Surface plasmon resonance (SPR) is an optical technique in which binding of an analyte to the surface changes the refractive index at the surface/solution interface. Molecular interactions are analysed in real time without a labeling step. Currently, the limit to SPR imaging is the small number of reactions that can be simultaneously analysed. Using a novel grafting technology and a new imaging system, we increased the throughput of SPR imaging. The interaction between p53 and DNA was chosen as a paradigm for validation of this assay. Using a tagged DNA methodology, we simultaneously targeted multiple DNA sequences on a single chip. The interaction between p53 and these DNA sequences was monitored by SPR imaging. Qualitative and quantitative analysis provides results similar to those obtained with conventional technologies.
Maillart
Emmanuel
E
Laboratoire Charles Fabry de l'Institut d'Optique (LCFIO), Centre National de la Recherche Scientifique CNRS UMR 8501, Bâtiment 503, Université Paris XI, 91403 Orsay, France.
Brengel-Pesce
Karen
K
Capela
Delphine
D
Roget
André
A
Livache
Thierry
T
Canva
Michael
M
Levy
Yves
Y
Soussi
Thierry
T
D004247:DNA / Q000378:metabolism*; D006801:Humans; D020349:Surface Plasmon Resonance / Q000379:methods*; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism*
D016159:Tumor Suppressor Protein p53; D004247:DNA