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Binding of [³H]Benzo(a)pyrene Metabolites to the AKR Mouse Embryo Cell Line Nuclear Proteins¹

Department of Cell Biology, Mayo Clinic, Rochester, Minnesota 55901

Nuclear proteins from AKR-2B mouse embryo cells incubated 24 hr with [³H]benzo(a)pyrene (1 µg/ml; 10 µCi/ml) were purified to remove nucleic acids and characterized by isoelectric focusing and sodium dodecyl sulfate gel electrophoresis on polyacrylamide gels. Analysis of the nuclear proteins by isoelectric focusing revealed a distribution of protein in the pH gradient with two prominent bands of protein, one focusing at pH 5 and the other at pH 11, both of which bound significant amounts of the carcinogen. The acid-soluble proteins from the nuclear preparation were analyzed by isoelectric focusing, and the results also revealed two peaks of protein and radioactivity at pH's of 5 and 11. Further analysis of the acid-soluble fraction on acid urea gels revealed that significant radioactivity was associated with proteins that migrated near but slower than histone H1 which may be the high-mobility group proteins 1 and 2. Radioactivity also comigrated with histones H3 and H2B. Low levels of radioactivity were associated with histone H1 and H4.

¹ Supported by Grant CA 22272 awarded by the National Cancer Institute, Department of Health, Education, and Welfare, the Mayo Foundation, and the Eagles Cancer Fund.

² To whom requests for reprints should be addressed.

Received 6/23/80. Accepted 1/22/81.