Membrane protein analysis using an improved peptic in-solution digestion protocol.
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.
Rietschel
Benjamin
B
Cluster of Excellence Macromolecular Complexes, Institute for Pharmaceutical Chemistry, Goethe-University, Frankfurt am Main, Germany.
Bornemann
Sandra
S
Arrey
Tabiwang N
TN
Baeumlisberger
Dominic
D
Karas
Michael
M
Meyer
Bjoern
B
D001436:Bacteriorhodopsins / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism; D019617:Halobacterium salinarum / Q000737:chemistry*; D010434:Pepsin A / Q000378:metabolism; D018717:Purple Membrane / Q000737:chemistry*; D012997:Solvents; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D053719:Tandem Mass Spectrometry
D012997:Solvents; D001436:Bacteriorhodopsins; D010434:Pepsin A