Abstract
Three electrophoretic techniques, isoelectric focusing, polyacrylamide electrophoresis and sodium dodecyl sulfate-polyacrylamide electrophoresis, were combined to develop a projective two-dimensional electrophoretic technique. The new technique was applied to analysis of human Cu, Zn-superoxide dismutase (SOD) isozymes. By using isoelectric focusing, human Cu, Zn-SOD resolved into five isozymes, which are thought to be charge isomers. From the result of the two-dimensional electrophoresis without denaturing agents, each isozyme was found to possess two or three electrophoretic forms, which were supposed to be in equilibrium. The five bands separated by electrophoresis thus contain several electrophoretic forms of SOD isozymes. The projective two-dimensional electrophoretic analysis indicates that all of the eleven electrophoretic forms consist of a single subunit with a molecular weight of 15.5kDa. These isozymes appear to be produced post-translationally.
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