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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Bacillus subtilis / chemistry
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Bacterial Proteins / analysis
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Bacterial Proteins / drug effects
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Electrophoresis, Polyacrylamide Gel
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Endopeptidase K / metabolism
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Glucose / pharmacology
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Humans
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Mass Spectrometry
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Membrane Proteins / analysis*
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Peptide Fragments / analysis*
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Peptide Hydrolases / metabolism*
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Proteomics / methods*
Substances
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Bacterial Proteins
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Membrane Proteins
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Peptide Fragments
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Peptide Hydrolases
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Endopeptidase K
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Glucose