Fractionation of chromosomal proteins of pea seedlings using procedures which avoid denaturation

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Chromatin was prepared from the buds and cotyledons of Alaska pea seedlings. The dissociated chromosomal components in the presence of 2 M NaCl and 5 M urea were completely fractionated into DNA and proteins with a Bio-Gel A50 column. The proteins were recovered by (NH_4)_2SO_4 and further fractionated into histones and non-histone proteins using a Bio-Rex 70(Na^+) column. The difference in the ratios of histones to non-histone proteins before and after chromatography with the Bio-Rex 70 was less than 10%. The histones and non-histone proteins thus prepared showed typical protein absorption spectra. Polyacrylamide gel electrophoresis of histones showed that the histone compositions in buds and cotyledon were similar, but the amount of H1 histones was a little less in cotyledons than in buds. Unlike histones, non-histone proteins fractionated by SDS-polyacrylamide gel electrophoresis indicated distinct differences between the two tissues. Buds had more heterogeneous non-histone proteins, at least 13 polypeptides, than cotyledons did. On the other hand, non-histone proteins of cotyledons showed less heterogeneity and lacked proteins of high molecular weight which were found in buds.
日本植物生理学会の論文