Mechanism of 67Ga accumulation in normal rat liver lysosomes.

67Ga accumulates in various malignant tumors and parenchymatous tissues. It was found to be associated with the soluble fraction of lysosomes (11). The present work investigates the mechanism of 67Ga accumulation in normal liver cells. Lysosomes were isolated from rat liver after intravenous injection of carrier free 67Ga. The soluble lysosomal fraction was obtained by sonication followed by centrifugation at 105,000 xg for 2 hrs. Gel filtration on Sephadex G 25 superfine was carried out on the soluble lysosomal fraction in order to investigate the stability of the 67Ga-protein complex within the lysosomes under EDTA treatment. After treatment with 1 mM/l EDTA a considerable amount of the protein bound radioactivity was found to be liberated. In further experiments the 67Ga binding lysosomal proteins were fractionated by electrophoresis on 7% polyacrylamide gels (0.5 cm x 5.5 cm). After staining with Coomassie blue 18 separated protein bands were apparent. 67Ga distribution within the gels was assessed by direct counting of radioactivity in gel slices. A considerable amount of the intralysosomal protein bound radioactivity migrated with a relative mobility of 0.36 corresponding to a protein band of molecular weight 85,000--90,000. This peak corresponded to the peak of 67Ga-labelled purified transferrin in control gels. These data were confirmed by immunoelectrophoresis combined with autoradiography: within the soluble lysosomal fraction a slight transferrin line could be identified. We conclude that 67Ga which is transported in the blood by transferrin (23) and taken up by the hepatic cell through endocytosis (32) is accumulated in the lysosomes associated with transferrin and its degraded fragments.