Evaluation of Endoproteinase Lys-C/Trypsin Sequential Digestion Used in Proteomics Sample Preparation
10.11895/j.issn.0253-3820.160801
- VernacularTitle:赖氨酸C端内切酶/胰蛋白酶顺序酶切在蛋白质组学样本制备中的评估
- Author:
Qian LI
;
Yu FENG
;
Minjia TAN
;
Linhui ZHAI
- Keywords:
Trypsin;
Lys-C;
Sequential digestion;
Proteomics;
Liquid chromatography-tandem mass spectrometry;
Missed cleavage
- From:
Chinese Journal of Analytical Chemistry
2017;45(3):316-321
- CountryChina
- Language:Chinese
-
Abstract:
Endoproteinase Lys-C/trypsin sequential digestion and trypsin digestion were used in 293 T 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.
