title: Phosphorylation impact on Spleen Tyrosine kinase conformation by Surface Enhanced Raman Spectroscopy.
abstract: Spleen Tyrosine Kinase (Syk) plays a crucial role in immune cell signalling and its altered expression or activation are involved in several cancers. Syk activity relies on its phosphorylation status and its multiple phosphorylation sites predict several Syk conformations. In this report, we characterized Syk structural changes according to its phosphorylation/activation status by Surface Enhanced Raman Spectroscopy (SERS). Unphosphorylated/inactive and phosphorylated/active Syk forms were produced into two expression systems with different phosphorylation capability. Syk forms were then analysed by SERS that was carried out in liquid condition on a lithographically designed gold nanocylinders array. Our study demonstrated that SERS signatures of the two Syk forms were drastically distinct, indicating structural modifications related to their phosphorylation status. By comparison with the atomic structure of the unphosphorylated Syk, the SERS peak assignments of the phosphorylated Syk nearest gold nanostructures revealed a differential interaction with the gold surface. We finally described a model for Syk conformational variations according to 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.
authors.affiliation: Université Paris 13, Sorbonne Paris Cité, Laboratoire CSPBAT, CNRS (UMR 7244), 74 rue Marcel Cachin, F-93017 Bobigny, France.|Université Paris 13, Sorbonne Paris Cité, Laboratoire ASIH, 74 rue Marcel Cachin, F-93017 Bobigny, France.|INSERM U978, Bobigny, France.
authors.forename: Maximilien
authors.initials: M
authors.lastname: Cottat
authors.affiliation: Université Paris 13, Sorbonne Paris Cité, Laboratoire CSPBAT, CNRS (UMR 7244), 74 rue Marcel Cachin, F-93017 Bobigny, France.
authors.forename: Ryohei
authors.initials: R
authors.lastname: Yasukuni
authors.affiliation: Université Paris 13, Sorbonne Paris Cité, Laboratoire CSPBAT, CNRS (UMR 7244), 74 rue Marcel Cachin, F-93017 Bobigny, France.|Université Paris 13, Sorbonne Paris Cité, Laboratoire ASIH, 74 rue Marcel Cachin, F-93017 Bobigny, France.|INSERM U978, Bobigny, France.
authors.forename: Yo
authors.initials: Y
authors.lastname: Homma
authors.affiliation: Université Paris 13, Sorbonne Paris Cité, Laboratoire CSPBAT, CNRS (UMR 7244), 74 rue Marcel Cachin, F-93017 Bobigny, France.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Lidgi-Guigui
authors.affiliation: Université Paris 13, Sorbonne Paris Cité, Laboratoire ASIH, 74 rue Marcel Cachin, F-93017 Bobigny, France.|INSERM U978, Bobigny, France.
authors.forename: Nadine
authors.initials: N
authors.lastname: Varin-Blank
authors.affiliation: Université Paris 13, Sorbonne Paris Cité, Laboratoire CSPBAT, CNRS (UMR 7244), 74 rue Marcel Cachin, F-93017 Bobigny, France.
authors.forename: Marc
authors.initials: M
authors.lastname: Lamy de la Chapelle
authors.affiliation: Université Paris 13, Sorbonne Paris Cité, Laboratoire ASIH, 74 rue Marcel Cachin, F-93017 Bobigny, France.|INSERM U978, Bobigny, France.
authors.forename: Christine
authors.initials: C
authors.lastname: Le Roy
mesh_terms: D000494:Allosteric Regulation; D000495:Allosteric Site / Q000235:genetics; D006801:Humans; D007107:Immune System / Q000502:physiology*; D010766:Phosphorylation; D011487:Protein Conformation; D015202:Protein Engineering; D015398:Signal Transduction; D013059:Spectrum Analysis, Raman / Q000379:methods; D000072377:Syk Kinase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: C000606896:SYK protein, human; D000072377:Syk Kinase