Studies on Protamines

XIII. The Fractionation of Clupeine Y

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1. More reproducible and satisfactory chromatographic systems on CM-cellulose (see Fig. 2) were found for fractionation of whole clupeine into Y and Z fractions in place of the previous method by adsorption column chromatography on alumina.
2. Further fractionation of clupeine Y was tried on a CM-cellulose column using several buffer systems over a wide pH range. Although YI rich and YII rich fractions were obtained, complete separa-tion of these 2 components could not be achieved in the case of intact clupeine Y.
3. From the chromatogram of clupeine Y fraction, however, clupeine Y was deduced to consist of nearly equal amounts of 2 main com-ponents, YI and YII, i.e. whole clupeine is composed of only 3 main species of molecules, YI, YII and Z. The amino acid compositions other than arginine of clupeine YI and YII were estimated as follows: 2Ala, 3Ser, 2Thr, 2Pro, lGly and llle for clupeine YI and 2Ala, 2Ser, lThr, 3Pro and 2Val for clupeine Yll.
4. In the case of whole clupeine modified with trinitrobenzenesul-fonic acid, all the 3 components of clupeine were successfully fractionated as TNP-YI, TNP-Z and free YII by one step elution from a CM-cel-lulose column.