Hepatic endosome fractions contain an ATP-driven proton pump.
Endosome fractions were isolated from rat liver homogenates on the basis of the subcellular distribution of circulating ligands, e.g. 125I-asialotransferrin internalized by hepatocytes by a receptor-mediated process. The distribution of endocytosed 125I-asialotransferrin 1-2 min and 15 min after uptake by liver and a monensin-activated Mg2+-dependent ATPase activity coincided on linear gradients of sucrose and Nycodenz. The monensin-activated Mg2+-ATPase was enriched relative to the liver homogenates up to 60-fold in specific activity in the endosome fractions. Contamination of the endosome fractions by lysosomes, endoplasmic reticulum, mitochondria, plasma membranes and Golgi-apparatus components was low. By use of 9-aminoacridine, a probe for pH gradients, the endosome vesicles were shown to acidify on addition of ATP. Acidification was reversed by addition of monensin. The results indicate that endosome fractions contain an ATP-driven proton pump. The ionophore-activated Mg2+-ATPase in combination with the presence of undegraded ligands in the endosome fractions emerge as linked markers for this new subcellular organelle.
Saermark
T
T
Flint
N
N
Evans
W H
WH
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000494:pharmacology*; D000585:Aminacrine / Q000378:metabolism; D000818:Animals; D001212:Asialoglycoproteins; D017301:Ca(2+) Mg(2+)-ATPase; D002259:Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone / Q000494:pharmacology; D002499:Centrifugation, Density Gradient; D066298:In Vitro Techniques; D008099:Liver / Q000187:drug effects / Q000378:metabolism*; D008985:Monensin / Q000494:pharmacology; D011522:Protons; D051381:Rats; D013347:Subcellular Fractions / Q000187:drug effects / Q000378:metabolism; D014168:Transferrin / Q000031:analogs & derivatives / Q000494:pharmacology
D001212:Asialoglycoproteins; D011522:Protons; D014168:Transferrin; C017031:asialotransferrins; D002259:Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone; D000585:Aminacrine; D000255:Adenosine Triphosphate; D008985:Monensin; D000251:Adenosine Triphosphatases; D017301:Ca(2+) Mg(2+)-ATPase