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Phosphorylation impact on Spleen Tyrosine kinase conformation by Surface Enhanced Raman Spectroscopy

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 charact...

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Autores principales: Cottat, Maximilien, Yasukuni, Ryohei, Homma, Yo, Lidgi-Guigui, Nathalie, Varin-Blank, Nadine, Lamy de la Chapelle, Marc, Le Roy, Christine
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5214100/
https://www.ncbi.nlm.nih.gov/pubmed/28054556
http://dx.doi.org/10.1038/srep39766
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author Cottat, Maximilien
Yasukuni, Ryohei
Homma, Yo
Lidgi-Guigui, Nathalie
Varin-Blank, Nadine
Lamy de la Chapelle, Marc
Le Roy, Christine
author_facet Cottat, Maximilien
Yasukuni, Ryohei
Homma, Yo
Lidgi-Guigui, Nathalie
Varin-Blank, Nadine
Lamy de la Chapelle, Marc
Le Roy, Christine
author_sort Cottat, Maximilien
collection PubMed
description 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.
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spelling pubmed-52141002017-01-09 Phosphorylation impact on Spleen Tyrosine kinase conformation by Surface Enhanced Raman Spectroscopy Cottat, Maximilien Yasukuni, Ryohei Homma, Yo Lidgi-Guigui, Nathalie Varin-Blank, Nadine Lamy de la Chapelle, Marc Le Roy, Christine Sci Rep Article 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. Nature Publishing Group 2017-01-05 /pmc/articles/PMC5214100/ /pubmed/28054556 http://dx.doi.org/10.1038/srep39766 Text en Copyright © 2017, The Author(s) http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Cottat, Maximilien
Yasukuni, Ryohei
Homma, Yo
Lidgi-Guigui, Nathalie
Varin-Blank, Nadine
Lamy de la Chapelle, Marc
Le Roy, Christine
Phosphorylation impact on Spleen Tyrosine kinase conformation by Surface Enhanced Raman Spectroscopy
title Phosphorylation impact on Spleen Tyrosine kinase conformation by Surface Enhanced Raman Spectroscopy
title_full Phosphorylation impact on Spleen Tyrosine kinase conformation by Surface Enhanced Raman Spectroscopy
title_fullStr Phosphorylation impact on Spleen Tyrosine kinase conformation by Surface Enhanced Raman Spectroscopy
title_full_unstemmed Phosphorylation impact on Spleen Tyrosine kinase conformation by Surface Enhanced Raman Spectroscopy
title_short Phosphorylation impact on Spleen Tyrosine kinase conformation by Surface Enhanced Raman Spectroscopy
title_sort phosphorylation impact on spleen tyrosine kinase conformation by surface enhanced raman spectroscopy
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5214100/
https://www.ncbi.nlm.nih.gov/pubmed/28054556
http://dx.doi.org/10.1038/srep39766
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