The effects of oxidation on the reverse-phase high-performance liquid chromatography characteristics of the high mobility groups 1 and 2 proteins.
Ion-pair reverse-phase high-performance liquid chromatography is a quick and convenient method for obtaining essentially pure preparations of the HMG (high mobility group)-1 and HMG-2 proteins if dithiothreitol is added to the eluted HMG protein fractions to prevent oxidation and their subsequent altered migration on acid-urea polyacrylamide gels. Unexpectedly, we found that this chromatographic separation technique can resolve the oxidized and reduced forms of both HMG-1 and HMG-2 proteins. We show that oxidized HMG-1 and -2 protein subfractions are responsible for some, but by no means all, of the HMG-1 and -2 protein heterogeneity previously reported by Elton and Reeves (2). At least two different HMG-1 protein species (one major and one minor) and at least four different HMG-2 protein species (two major and two minor) are consistently found in fully reduced "enriched" HMG-1 and -2 pig thymus protein preparations.
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D006609:High Mobility Group Proteins / Q000302:isolation & purification*; D010084:Oxidation-Reduction; D013552:Swine; D013950:Thymus Gland / Q000032:analysis
D006609:High Mobility Group Proteins