Univerzita Pardubice Fakulta chemicko- technologická

Univerzita Pardubice

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English

Publikace detail

Optimization of sample preparation method for the analysis of protein phosphorylation by MALDI-MS and CZE

Autoři: Staněk Václav | Jankovičová Barbora | Kupčík Rudolf | Bennet Aneta | Coufalová Eva | Bílková Zuzana

Rok: 2015

Druh publikace: ostatní - přednáška nebo poster

Strana od-do: nestránkováno

Tituly:

Jazyk	Název	Abstrakt	Klíčová slova
eng	Optimization of sample preparation method for the analysis of protein phosphorylation by MALDI-MS and CZE	This work is focused on optimization of sample preparation method for the analysis of phosphopeptides using MALDI LTQ Orbitrap mass spectrometry and capillary zone electrophoresis. All sample preparation steps were tested on bovine α-casein, which was chosen as a model phosphoprotein. Sample of α-casein was digested by the proteolytic enzyme trypsin, immobilized on two different types of magnetic microparticles, macroporous pearl cellulose (Perloza MG) and magnetic silica beads SiMAG-COOH. Phosphopeptides occurring in the peptide sample were enriched using titanium dioxide particles and dephosphorylated by alkaline phosphatase (ALP), immobilized on Perloza MG and SiMAG-NH2 magnetic microparticles. The highest activity of ALP immobilized on SiMAG-NH2 microparticles was achieved in Tris-HCl buffer, while maximal activity of ALP immobilized on Perloza MG microparticles was evaluated using phosphate buffer. All adjustment steps of model sample were verified by MALDI-MS prior to CZE analysis.	MALDI-MS, CZE, protein phosphorylation, phosphopeptides