A lysosome- and mitochondria-enriched fraction was obtained from rat skeletal muscle using a differential centrifugation procedure. This fraction contained a proportion (3-4%) of the tissue glycogen content. 2. Lysosomes and mitochondria were separated from one another by centrifugation of a cell-free extract upon discontinuous Ficoll-sucrose gradients. Little glycogen was associated with the resulting mitochondrial fraction. The lysosomal fraction, however, contained a significant amount of glycogen, accounting for 5% of the skeletal muscle glycogen. 3. The lysosomal glycogen was purified and found to be enriched in high molecular weight material. 4. The compartmentation of skeletal muscle glycogen metabolism is suggested.
