Endocytosis and breakdown of 125I-labelled lactate dehydrogenase isoenzyme M4 by rat liver and spleen in vivo.

1. Porcine lactate dehydrogenase isoenzyme M4 was labelled with 125I and injected intravenously into rats. Enzyme activity and radioactivity in plasma were cleared in an identical way with a half-life of about 30 min. This half-life was the same as that of unlabelled enzyme. 2. Uptake of label by liver and spleen was determined. Radioactivity in these tissues increased up to about 13 min after injection and subsequently declined. Radioautography indicated uptake of the enzyme by sinusoidal liver cells (probably Kupffer cells) and by spleen macrophages. After differential fractionation of liver, acid-precipitable radioactivity was largely found in the light mitochondrial and microsomal fractions, suggesting localization in lysosomes and endosomes respectively. 3. The amount of acid-soluble radioactive breakdown products in plasma started to rise between 7 and 15 min after injection. Breakdown in liver and spleen was retarded by previous injection of suramin, an inhibitor of lysosomal proteolysis. 4. The contribution of liver and spleen towards the clearance of the enzyme could be calculated from its half-life in plasma and its uptake by the organs within the first 13 min period after injection. Our results indicate that about 65% and 12% of the injected dose was taken up, and subsequently broken down, by liver and spleen respectively. 5. Unlabelled porcine lactate dehydrogenase isoenzyme H4 showed a plasma half-life of about 8 h. This isoenzyme is therefore endocytosed by liver at a much slower rate than isoenzyme M4 (if it is taken up at all).