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Fractionation of -Casein on a Diethylaminoethyl Cellulose Column¹

Division of Applied Biology, National Research Council, Ottawa, Canada

ABSTRACT

A complex chromatogram showing seven overlapping peaks was obtained from -casein by gradient sodium chloride elution (0 0.4 M), followed by 0.25 N sodium hydroxide, from a diethylaminoethyl cellulose column. The material recovered from the two peaks eluted most quickly contained no -casein, but contained several components with low mobility in starch gel electrophoresis (SGE), and low sialic acid and phosphate contents. A later fraction contained a high proportion of a casein resembling _s-casein, with -casein and some components that moved more rapidly than _s-casein in SGE.

Recoveries from the four other peaks were mainly -caseins that differed in their elution characteristics but not in SGE patterns, sialic acid content, or phosphate content. One was not eluted with sodium chloride but was eluted in advance of the sodium hydroxide front; another was eluted with the sodium hydroxide front. Both latter peaks may represent an aggregated form of -casein.

Chromatography achieved only limited purification and was accompanied by a deleterious aging of the -casein. This aging released a sialic acid-containing compound soluble in 12% trichloracetic acid, and lowered the ability of the -casein to stabilize _s-casein in the presence of calcium.

¹ Issued as N.R.C. No. 7732.