Author(s): PAN Li-mei, WEI Shu-gen, FU Qiang, TAN Yun, ZHU Jia-jie, Guangxi Botanical Garden of Medicinal Plant, State Key Laboratory of Conservation and Utilization of Subtropical Agro-bioresource, Guangxi University, Guangxi Fisheries Research Institute
Pages: 612-615+619
Year: 2015 Issue:  6
Journal: Guangxi Sciences
Keyword:  in-solution enzymatic digestion; proteins; LC-MS; peptides coverage;
Abstract: 【Objective】Two methods of proteins in-solution enzymatic digestion were compared and evaluated to establish a reliable sample preparation protocol for large-scale identification in proteomics research.【Methods】Bovine serum albumin(BSA)was reduced and alkylated,and subsequently used for in-solution enzymatic digestion according to the protocols of group A(TEAB method)and group B(TFE method).LTQ-Orbitrap mass spectrometer was utilized to obtaine the signal of enzymatic peptides.The data obtained from two groups were compared.Peptide categories,variable modification site and protein coverage were analyzed.【Results】A total of 36 peptides and 39 modification sites were acquired in group A(TEAB method),and the protein coverage was 59.31%;in group B(TFE method),a total of 27 peptides and 27 modification sites were obtained,and the protein coverage was 47.83%.All the statistical compari-sons between groups A and B were significantly different.【Conclusion】TEAB method provided a high efficiency of enzymatic digestion with higher peptide coverage.This method can be served as the experimental basis for MS analysis in proteomics.