University of PardubiceFaculty of Chemical Technology

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An improved detection of multiphosphorylated peptides using novel magnetic TiO2 based nanomaterial

Authors: Kupčík Rudolf | Řehulka Pavel | Macák Jan | Fabrik Ivo | Klimentová Jana | Řehulková Helena | Murasová Pavla | Stulík Jiří | Bílková Zuzana

Year: 2017

Type of publication: ostatní - přednáška nebo poster

Page from-to: nestránkováno

Titles:

Language	Name	Abstract	Keywords
eng	An improved detection of multiphosphorylated peptides using novel magnetic TiO2 based nanomaterial	he aim of our work is to demonstrate the magnetic nanomaterial based on TiO2 with nanotubular structural state which increases the number of eluted and analyzed multiphosphorylated peptides, as compared to common TiO2 materials available today. A peptide mixture consisted of tryptically digested phosphorylated and nonphosphorylated proteins was used as simple model mixture. The phosphopeptide enrichment was performed either with TiO2 microparticles (Titansphere, GL Sciences) or with the novel magnetic nanomaterial. Binding was done in 80% acetonitrile containing trifluoroacetic and lactic acid and elution was done using 1% ammonia solution. Mass spectrometry analysis was done using Thermo Scientific MALDI LTQ Orbitrap XL mass spectrometer. Also a complex peptide mixture derived from stimulated Jurkat cells was processed using the similar protocol and analyzed by LC-MS/MS (Thermo Scientific UltiMate3000 RSLC-nanoHPLC system and Q Exactive mass spectrometer). Proteome Discoverer software (Thermo Scientific, v. 2.0) was used for identification of MS/MS spectra and R software was used for data processing. The presented magnetic TiO2 nanotubular nanomaterial showed the high enrichment efficiency both for monophosphorylated and multiphosphorylated peptides that are generally difficult to elute from TiO2. Presented results indicate the improved performance in the analysis of multiphosphorylated peptides, which is caused by a different structural design of the presented material as compared to common TiO2 based stationary phases. These properties predetermine this material as useful tool for phosphoproteomic studies. The analysis of phosphopeptides can be thus carried out in a single enrichment step as compared to SIMAC protocol (1) while still using the advantageous isolation selectivity. Next important advantage of the presented material is its magnetic properties providing an easier handling during all enrichment steps.	Multiphosphorylated peptides; magnetic nanomaterial; TiO2