摘要
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.
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.
基金
Supported by the National Natural Science Foundation of China（Nos.81472454, 31100930）, the Natural Science Foundation of Jilin Provincial Science and Technology Department, China（Nos.2011713, 20090461, 20110739, 20130413017GH）, the Natural Science Foundation of Heilongjiang Province, China（Nos.201000742, D201075, Gc09c317）, the Natural Science Foundation of Jilin Provincial Development and Reform Commission, China（No.2013c026-5） and the Natural Science Foundation of Changchun Science and Technology Bureau, China（No.2011125）.
