A procedure had been devised by which
in vivo liver catalase-depressing substance is extractable as its aqueous solution from particles of the cells of normal muscle and Rhodamine sarcoma of rats.3) Some nature of the solubilized substance was studied.
1) By molecular sieve fractionation of the extract, either on Sephadex G-75 column or on Sephadex G-200 column, the
in vivo catalase-depressing activity of the extract was separated into two fractions, excluded and diffused, indicating that the extract contained at least two kinds of active substance different in molecular size.
2) When the extract was dialyzed and then adjusted to pH 4.0, precipitation occurred. Both precipitate and supernatant showed the activity. By the isoelectric fractionation with Ampholine-carrier Ampholyte, the supernantant was divided into fractions of different isoelectric points of 3.39-3.43, 3.44-3.90, 4.05-4.60, 4.82-5.31, 5.45-6.75, 7.19-8.76, and 8.82-9.48. All the fractions but that of pI 7.19-8.76 showed the activity, indicating that the supernatant contained various active substances different in isoelectric point.
From these findings, it is concluded that the
in vivo catalase-depressing substance varies in chemical composition and/or in mode of existence.
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