The effect of isoproterenol on nuclear protein synthesis in electrostatically sorted rat hepatocytes

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Abstract
Rat hepatocyte nuclei were electrostatically sorted and collected from specific regions of the cell cycle. Proteins were extracted from homogeneous populations of nuclei with cetyltrimethylammonium bromide and chromatographed by one- and two-dimensional acid-urea polyacrylamide gel electrophoresis. Small quantitative differences were observed in amido black stained two-dimensional patterns of the nuclear proteins of the G0 + G1 and G2 + M populations. However, comparison of the two-dimensional autoradiographs of 3H-leucine labeled nuclear proteins in these populations revealed dramatic differences. After isoproterenol administration, animals showed enhanced labeling of certain nuclear proteins in G2 + M populations, but no noticeable effect in protein labeling was observed in the G0 + G1 populations. Four days after the administration of isoproterenol, fluorescent histograms exhibited increased amounts of DNA in the G2 + M populations. Comparison of the total amino acid composition of two proteins from control animals revealed the distinct chemical contrast of these proteins. Both proteins possessed higher ratios of acidic to basic residues than anticipated for basic proteins. These procedures and findings should prove useful for further understanding of mechanisms of toxicant effects upon nuclear protein metabolism.