Citation: SUN Li, ZHAO Hai-Feng, GUO Hong-Hua, YANG Guang-Yuan, HE Cheng-Yan, SHI Qing-Hong, WEN Xue, WANG Xiao-Ting, ZHAO Li-Chun. Mass Spectrometric Analysis of Phosphorylation Modification in Talin from Human Colorectal Carcinoma Tissues[J]. Chinese Journal of Analytical Chemistry, ;2012, 40(10): 1500-1506. doi: 10.3724/SP.J.1096.2012.20027 shu

Mass Spectrometric Analysis of Phosphorylation Modification in Talin from Human Colorectal Carcinoma Tissues

1.
¹ China-Japan Union Hospital, Jilin University, Changchun 130033, China;

Corresponding author: ZHAO Li-Chun,
Received Date: 10 January 2012
Available Online: 21 March 2012
Fund Project: 本文是国家自然科学基金(No.21175055) (No.21175055) 吉林省科技厅项目(Nos.2011713,20110739)资助 (Nos.2011713,20110739)

The phosphorylation modification of talin, especially the phosphorylation state of talin in pathological environment such as carcinoma, is closely relevant to carcinogenesis and metastasis process. In this study, talin protein was isolated from human colorectal carcinoma tissues by salt fractionation and ion exchange chromatography, followed by further purification by electrophoresis. The purified talin was subject to tryptic digestion. Either immobilized Fe³⁺ affinity chromatography or TiO₂ affinity chromatography was used to absorb the phosphorylated peptides under acidic condition, which were then eluted with 1% ammonium hydroxide. The separation was performed on a Michrom Magic C₁₈ column with gradient elution using two mobile phase solutions (A:99% water+1% ACN+0.1% formic acid; B:99% ACN+1% water+0.1% formic acid). ESI mass spectrometer was used to detect the product ions of the eluted peaks under data-dependent acquisition mode. The results indicated that 8 phosphorylated peptides were captured by the immobilized Fe³⁺ affinity chromatography, whereas 9 phosphorylated peptides were captured by TiO₂ affinity chromatography. The present study provides a rapid, accurate method for characterizing talin protein isolated from human colorectal carcinoma tissues.
- Mass spectrometry,
- Talin,
- Colorectal carcinoma,
- Phosphorylation
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