Uptake of tyramine cellobiose by rat liver.
The uptake of 125I-tyramine cellobiose (TC) by isolated rat hepatocytes and by total rat liver is markedly higher than that of 14C-sucrose and 125I-PVP, suggesting that TC does not enter the cells by fluid phase endocytosis. The distribution of radioactivity after differential centrifugation shows that the compound is shared out amongst sedimentable structures and unsedimentable fraction. Analysis by isopycnic centrifugation indicates that quickly after its penetration into the cells, most of sedimentable 125I-TC is associated with lysosomes. Such an intracellular localization is confirmed by the distributions observed after free flow electrophoresis and by the fact that radioactivity and cathepsin C, a lysosomal hydrolase, are simultaneously released from a mitochondrial fraction treated with glycyl-L-phenylalanine-2-naphthylamide. Pretreatment of the rats with chloroquine, an acidotropic drug that accumulates in lysosomes, prevents to some extent the entry of 125I-TC into these organelles. Experiments performed with purified lysosomes show that 14C-sucrose does not cross the lysosomal membrane when 125I-TC accumulates linearly with time in the fractions. These results are explained by supposing that the linkage of tyramine to cellobiose allow the disaccharide to diffuse through the plasma and the lysosome membranes, and that the accumulation of the molecule in these organelles results from its weak basic properties. 125I-TC could be an interesting molecule with which to study acidotropism in the whole animal and in isolated and cultured cells.
Zhong
Z D
ZD
Laboratoire de Chimie Physiologique, Facultés Universitaires Notre-Dame de la Paix, Namur, Belgium.
Jadot
M
M
Wattiaux-De Coninck
S
S
Wattiaux
R
R
D000818:Animals; D001692:Biological Transport; D002250:Carbon Radioisotopes; D002475:Cellobiose / Q000378:metabolism*; D002478:Cells, Cultured; D007457:Iodine Radioisotopes; D007700:Kinetics; D008099:Liver / Q000378:metabolism*; D008297:Male; D011206:Povidone-Iodine / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D013395:Sucrose / Q000378:metabolism; D014439:Tyramine / Q000378:metabolism*
D002250:Carbon Radioisotopes; D007457:Iodine Radioisotopes; C038394:tyramine-cellobiose adduct; D002475:Cellobiose; D013395:Sucrose; D011206:Povidone-Iodine; D014439:Tyramine