Application of Chromatography to the Separation of Subcellular, Enzymatically Active Granules

Abstract

Purification and concn. of melanized granules from saline extract of Cloudman S91 and Harding-Passey mouse melanomas is accomplished by adsorption on No. 503 Celite column, development of column with 0.154 [image] NaCl, extrusion and partition of column, and elution with H2O or NaCl soln. Elution may be accomplished by flowing chromatography without extrusion. Chromatographed granules have higher ratio of succinoxidase and dopa-oxidase activity per mg. of N than beginning extracts by several hundred percent, and the ratio is highest for the first part of the column. Concn. of granules in elutrate may be increased by centrifuga-tion. No changes in morphology or staining reactions of granules result from this treatment. An electron micrograph of the chromatographed granules is presented.