Proteomics of mouse liver microsomes: performance of different protein separation workflows for LC-MS/MS.
The mouse liver microsome proteome was investigated using ion trap MS combined with three separation workflows including SDS-PAGE followed by reverse-phase LC of in-gel protein digestions (519 proteins identified); 2-D LC of protein digestion (1410 proteins); whole protein separation on mRP heat-stable column followed by 2-D LC of protein digestions from each fraction (3-D LC; 3703 proteins). The higher number of proteins identified in the workflow corresponded to the lesser percentage of run-to-run reproducibility. Gel-based method yielded a number of predicted membrane proteins similar to LC-based workflows.
Zgoda
Victor G
VG
Institute of Biomedical Chemistry, Moscow, 119121, Russia.
Moshkovskii
Sergei A
SA
Ponomarenko
Elena A
EA
Andreewski
Timofey V
TV
Kopylov
Arthur T
AT
Tikhonova
Olga V
OV
Melnik
Stanislav A
SA
Lisitsa
Andrei V
AV
Archakov
Alexander I
AI
D000818:Animals; D002853:Chromatography, Liquid; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D008297:Male; D051379:Mice; D008862:Microsomes, Liver / Q000378:metabolism*; D040901:Proteomics; D053719:Tandem Mass Spectrometry