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.
Bendz
Maria
M
Protein Technology, Department of Immunotechnology, CREATE Health, Lund University, Sweden.
Möller
Mirja Carlsson
MC
Arrigoni
Giorgio
G
Wåhlander
Asa
A
Stella
Roberto
R
Cappadona
Salvatore
S
Levander
Fredrik
F
Hederstedt
Lars
L
James
Peter
P
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