[16p-F206-1~19] 12.6 Nanobiotechnology

[16p-F206-19] Phosphorylation impact on protein conformation by Surface Enhanced Raman Spectroscopy

〇Ryohei Yasukuni^1,2, Cottat Maximilien^2,3, Homma Yo², Lidgi-Guigui Nathalie², Varin-Blank Nadine^2,3, Lamy de la Chapelle Marc², Le Roy Christine^2,3 (1.NAIST, 2.Univ. Paris 13, 3.INSERM)

Keywords:Surface Enhanced Raman Spectroscopy, Protein conformation, Biosensor

We characterized protein structural changes associated with phosphorylation status by Surface Enhanced Raman Spectroscopy (SERS). As a sample protein, Unphosphorylated and phosphorylated forms of Spleen Tyrosine Kinase (Syk) were produced into two expression systems. Syk forms were then analyzed by SERS that was carried out in liquid condition on a lithographically designed gold nanocylinders array. The obtained SERS spectra of the two Syk forms were drastically distinct, indicating structural modifications related to their phosphorylation status. The SERS peaks were assigned on the basis of differential interactions with the gold surface using known atomic structure of the unphosphorylated Syk. We finally describe a model for Syk conformational variations involved in its phosphorylation status. In conclusion, SERS is an efficient technical approach for studying in vitro protein conformational changes and might be a powerful tool to determine protein functions in tumour cells.