1. Proteomics of mouse liver microsomes: performance of different protein separation workflows for LC-MS/MS
- Author
-
Andrei V. Lisitsa, Alexander I. Archakov, O. V. Tikhonova, Victor G. Zgoda, Sergei A. Moshkovskii, Arthur T. Kopylov, Stanislav A. Melnik, Timofey V. Andreewski, and Elena A. Ponomarenko
- Subjects
Male ,Proteomics ,Chromatography ,Protein digestion ,Complete protein ,Biology ,Biochemistry ,Mice ,Membrane protein ,Tandem Mass Spectrometry ,Proteome ,Protein purification ,Microsome ,Microsomes, Liver ,Animals ,Electrophoresis, Gel, Two-Dimensional ,Electrophoresis, Polyacrylamide Gel ,Ion trap ,Molecular Biology ,Chromatography, Liquid - Abstract
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.
- Published
- 2009