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Citation: LI Qian, FENG YU, TAN Min-Jia, ZHAI Lin-Hui. Evaluation of Endoproteinase Lys-C/Trypsin Sequential Digestion Used in Proteomics Sample Preparation[J]. Chinese Journal of Analytical Chemistry, ;2017, 45(3): 316-321. doi: 10.11895/j.issn.0253-3820.160801 shu

Evaluation of Endoproteinase Lys-C/Trypsin Sequential Digestion Used in Proteomics Sample Preparation

LI Qian ^1,2, ,
FENG YU ³ ,
TAN Min-Jia ^1,2 ,
ZHAI Lin-Hui ¹

1.
State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China;
2.
University of Chinese Academy of Sciences, Beijing 100049, China;
3.
Beijing Hualishi Technology Co., Ltd., Beijing 101312, China

Received Date: 2 November 2016
Revised Date: 23 December 2016

Fund Project: This work was supported by Youth Science and Technology Talents in Shanghai Sailplan (No. 16YF141400) the National Natural Science Foundation of China (No.31370814) and Shanghai Science and Technology Committee (No.14DZ2261100)

Endoproteinase Lys-C/trypsin sequential digestion and trypsin digestion were used in 293T cell proteomics sample preparation and the results of Lys-C/trypsin sequential digestion and trypsin digestion in proteomics sample preparation was systematically evaluated. It was found that the number of identified peptides and proteins increased significantly, and missed cleavage sites, especially K sites decreased dramatically through Lys-C/trypsin sequential digestion. And the average sequence coverage of identified proteins in Lys-C/trypsin sequential digestion sample was higher than that in trypsin digestion sample. Besides, different amount of enzymes was tested to select the optimal usage of enzymes in Lys-C/trypsin sequential digestion. This study provided the references for proteomics sample preparation.
- Trypsin,
- Lys-C,
- Sequential digestion,
- Proteomics,
- Liquid chromatography-tandem mass spectrometry,
- Missed cleavage
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