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Citation: HONG Xin, QIAO Lu, YANG Jing-Bo, SUN Xiao-Ying, WANG Hai, HE Cheng-Yan. Mass Spectrometric Analysis of Proteoforms of Cyclophilin A from Human Colorectal Carcinoma Tissues[J]. Chinese Journal of Analytical Chemistry, ;2018, 46(2): 195-202. doi: 10.11895/j.issn.0253-3820.171393 shu

Mass Spectrometric Analysis of Proteoforms of Cyclophilin A from Human Colorectal Carcinoma Tissues

1.
Department of Vascular Surgery, China-Japan Union Hospital, Jilin University, Changchun 130033, China;
2.
Clinical Laboratory, China-Japan Union Hospital, Jilin University, Changchun 130033, China;
3.
Central Laboratory, Second Hospital, Jilin University, Changchun 130014, China

Received Date: 6 November 2017
Revised Date: 14 December 2017

Fund Project: This work was supported by the National Natural Science Foundation of China (No. 81572082) and the Jilin Province Science and Technology Department (Nos. 20150414015GH, 2011713).

Cyclophilin A was isolated from human colorectal carcinoma tissues by ultrafiltration, affinity chromatography, and two dimensional electrophoresis. By the aid of Western blot analysis, several isoforms of cyclophilin A were detected., The tryptic digests from cyclophilin A were analyzed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry as well as electrospray tandem mass spectrometry techniques. Several different modifications such as N-terminal acetylation (Ac-¹Met and Ac-²Val) and phosphorylation of ⁵Thr were identified from these isoforms of cyclophilin A, which represented the majority of the proteoforms of cyclophilin A in this study. In addition, other kinds of modifications such as oxidation and deamidation were also found in these proteoforms. The results indicated that the developed method could be used to rapidly and correctly identify the several proteoforms of cyclophilin A isolated from human colorectal carcinoma tissues.
- Mass spectrometry,
- Cyclophilin A,
- Colorectal carcinoma,
- Acetylation,
- Phosphorylation
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