Membrane protein analysis using an improved peptic in-solution digestion protocol

Proteomics. 2009 Dec;9(24):5553-7. doi: 10.1002/pmic.200900532.

Abstract

In the proteomic analysis of membrane proteins, less-specific proteases have become a promising tool to overcome fundamental limitations of trypsin with its unique specificity for basic residues. Pepsin is well-known to be utilized for specific applications that require acidic conditions, but in terms of membrane protein identification and characterization, it has been disregarded for the most part. This work presents an optimization of an existing peptic digest protocol for the analysis of membrane proteins using bacteriorhodopsin from purple membranes as reference.

MeSH terms

  • Bacteriorhodopsins / analysis*
  • Bacteriorhodopsins / isolation & purification
  • Bacteriorhodopsins / metabolism
  • Halobacterium salinarum / chemistry*
  • Pepsin A / metabolism
  • Purple Membrane / chemistry*
  • Solvents
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Tandem Mass Spectrometry

Substances

  • Solvents
  • Bacteriorhodopsins
  • Pepsin A