| Abstract: | Actin was extracted from isolated nuclei of Acanthamoeba castellanii and purified to homogeneity under nondenaturing conditions by diethylaminoethylcellulose and Sephadex G-100 chromatography. The pure protein has the same molecular weight as cytoplasmic Acanthamoeba actin and a very similar amino acid composition. Isoelectrofocusing shows that nuclear actin is slightly more acidic than the major cytoplasmic species, and comparative analysis of peptides from tryptic and cyanogen bromide digests shows that both actins are very similar but not chemically identical. In an assay that is specific for most actins, the inhibition of DNase I through the formation of a 1:1 G-actin-DNase I complex, the nuclear and cytoplasmic actins are equally effective. By use of a similar procedure for the purification of both actins, it is estimated that the amount of nuclear actin is about 1.5% of the amount of cytoplasmic actin, a major protein of the amoeba. It is concluded that a minor isoelectric species of actin associates selectively with the nuclei of A. castellanii. |
