A new insight into the impact of different proteases on SILAC quantitative proteome of the mouse liver

Proteomics. 2013 Aug;13(15):2238-42. doi: 10.1002/pmic.201200590. Epub 2013 Jun 20.

Abstract

In this study, we examined the use of multiple proteases (trypsin, LysC, tandem LysC/trypsin) on both protein identification and quantification in the Lys-labeled SILAC mouse liver. Our results show that trypsin and tandem LysC/trypsin digestion are superior to LysC in peptides and protein identification while LysC shows advantages in quantification of Lys-labeled proteins. Combination of experimental results from different proteases (LysC and trypsin) enabled a significant increase in the number of identified protein and protein can be quantified. Thus, taking advantage of the complementation of different protease should be a good strategy to improve both qualitative and quantitative proteomics research.

Keywords: LysC; Quantification; SILAC mouse liver; Technology; Trypsin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Isotope Labeling / methods*
  • Liver / chemistry*
  • Liver / metabolism
  • Metalloendopeptidases / metabolism*
  • Mice
  • Peptide Fragments / analysis*
  • Peptide Fragments / chemistry
  • Peptide Fragments / metabolism
  • Proteins / analysis
  • Proteins / chemistry
  • Proteins / metabolism
  • Proteome / analysis*
  • Proteome / chemistry
  • Proteome / metabolism
  • Proteomics / methods
  • Tandem Mass Spectrometry
  • Trypsin / metabolism*

Substances

  • Peptide Fragments
  • Proteins
  • Proteome
  • Trypsin
  • Metalloendopeptidases
  • peptidyl-Lys metalloendopeptidase