Mass spectrometric analysis revealing phosphorylation modifications of periostin

Abstract

Human periostin was over-expressed in HEK293T cells, which was enriched by nickel ion affinity resin, and further purified by gel electrophoresis. Phosphopeptides contained in the tryptic digestion of the purified periostin were enriched by TiO₂ affinity chromatography, and then analyzed by liquid chromatography-tandem mass spectrometry(LC-MS/MS). LC-MS/MS analysis reveals three phosphorylation modification sites of periostin at IKVIEGpSLQPIIK(682–694), pSLHEKLKQDK(498–507) and p[TT]VLYEC*C*PGYM*R(73–85). The established method could be a great help to profiling the phosphorylation status of periostin under different physiological environments, such as inflammatory and tumor micro-environments.

Keywords

Mass spectrometry / Periostin / Phosphorylation

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