Quantification of membrane proteins using nonspecific protease digestions.
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.
Protein Technology, Department of Immunotechnology, CREATE Health, Lund University, Sweden.
Maria
M
Bendz
Mirja Carlsson
MC
Möller
Giorgio
G
Arrigoni
Asa
A
Wåhlander
Roberto
R
Stella
Salvatore
S
Cappadona
Fredrik
F
Levander
Lars
L
Hederstedt
Peter
P
James
D001412:Bacillus subtilis / Q000737:chemistry; D001426:Bacterial Proteins / Q000032:analysis / Q000187:drug effects; D004591:Electrophoresis, Polyacrylamide Gel; D019286:Endopeptidase K / Q000378:metabolism; D005947:Glucose / Q000494:pharmacology; D006801:Humans; D013058:Mass Spectrometry; D008565:Membrane Proteins / Q000032:analysis*; D010446:Peptide Fragments / Q000032:analysis*; D010447:Peptide Hydrolases / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*
D001426:Bacterial Proteins; D008565:Membrane Proteins; D010446:Peptide Fragments; D010447:Peptide Hydrolases; D019286:Endopeptidase K; D005947:Glucose