Studies on unknown methylated compounds of non-histone nuclear protein

Abstract

The HCl hydrolyzate of the non-histone protein fractionated from the rat liver nuclei which have been incubated in the presence of S-adenosyl-l-[methyl-¹⁴C]-methionine shows at least four unidentified radioactive peaks on a basic amino acid analysis chromatogram. One of these unknown compounds (designated as compound 3) is also formed by the rat liver homogenate with the exogenous addition of an appropriate protein substrate. Since boiled rat liver homogenate or fresh homogenate in the absence of an exogenous protein substrate failed to form compound 3, its formation can be considered to be enzyme-catalyzed. The enzyme which yields compound 3 shows a preference of protein substrate in the order of reductively methylated hemoglobin >native >histone type II-A. The rat enzyme is nuclear in location associated with chromatin, and exhibits the highest activity in the liver among various rat organs. A compound 3-forming enzyme is also present inNeurospora crassa, since endogenous formation of the compound 3 can be demonstrated with the crude extract of this mold. The chemical identity of compound 3 is not yet known. However, it resisted to the following treatments; 6 N HCl and 1.0 N NaOH hydrolysis at 110°C, orl-amino acid oxidase.