Quantification of Membrane Proteins Using Nonspecific Protease Digestions

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Abstract

We present a mass spectrometry-based method for the identification and quantification of membrane proteins using the low-specificity protease Proteinase K, at very high pH, to digest proteins isolated by a modified SDS-PAGE protocol. The resulting peptides are modified with a fragmentation-directing isotope labeled tag. We apply the method to quantify differences in membrane protein expression of Bacillus subtilis grown in the presence or absence of glucose.

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Research areas and keywords

Subject classification (UKÄ) – MANDATORY

  • Immunology in the medical area

Keywords

  • membrane proteins, N-terminal labeling, quantification, relative, proteinase K, Bacillus subtilis
Original languageEnglish
Pages (from-to)5666-5673
JournalJournal of Proteome Research
Volume8
Issue number12
Publication statusPublished - 2009
Publication categoryResearch
Peer-reviewedYes