We performed a threedimensional separation of pulsechase duallabelled rat liver cytosolic proteins using hydrophobic interaction chromatography, isoelectric focusing, and SDS gel electrophoresis. Due to very different expression rates but similar size and pI of rat liver cytosolic proteins, we demonstrate the impossibility of successful twodimensional separations of such complex protein mixtures. A prefractionation of proteins by hydrophobic interaction chromatography is therefore recommended prior to twodimensional gel electrophoresis. Our studies confirmed the correlation between protein turnover rates and surface hydrophobicity.
Print ISSN: 1431-6730
Volume: 382, 04/2001
Pages: 677 - 682