Dehydroascorbate and ascorbate transport in rat liver microsomal vesicles.
Ascorbate and dehydroascorbate transport was investigated in rat liver microsomal vesicles using radiolabeled compounds and a rapid filtration method. The uptake of both compounds was time- and temperature-dependent, and saturable. Ascorbate uptake did not reach complete equilibrium, it had low affinity and high capacity. Ascorbate influx could not be inhibited by glucose, dehydroascorbate, or glucose transport inhibitors (phloretin, cytochalasin B) but it was reduced by the anion transport inhibitor 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid and by the alkylating agent N-ethylmaleimide. Ascorbate uptake could be stimulated by ferric iron and could be diminished by reducing agents (dithiothreitol, reduced glutathione). In contrast, dehydroascorbate uptake exceeded the level of passive equilibrium, it had high affinity and low capacity. Glucose cis inhibited and trans stimulated the uptake. Glucose transport inhibitors were also effective. The presence of intravesicular reducing compounds increased, while extravesicular reducing environment decreased dehydroascorbate influx. Our results suggest that dehydroascorbate transport is preferred in hepatic endoplasmic reticulum and it is mediated by a GLUT-type transporter. The intravesicular reduction of dehydroascorbate leads to the accumulation of ascorbate and contributes to the low intraluminal reduced/oxidized glutathione ratio.
Bánhegyi
G
G
Istituto di Patologia Generale, Università di Siena, 53100 Siena, Italy. banhegyi@puskin.sote.hu
Marcolongo
P
P
Puskás
F
F
Fulceri
R
R
Mandl
J
J
Benedetti
A
A
D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid / Q000494:pharmacology; D000818:Animals; D001205:Ascorbic Acid / Q000378:metabolism*; D001692:Biological Transport / Q000187:drug effects; D002712:Chlorides; D003683:Dehydroascorbic Acid / Q000378:metabolism*; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D005033:Ethylmaleimide / Q000494:pharmacology; D005290:Ferric Compounds / Q000494:pharmacology; D005947:Glucose / Q000494:pharmacology; D008297:Male; D008862:Microsomes, Liver / Q000378:metabolism*; D009004:Monosaccharide Transport Proteins / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013347:Subcellular Fractions / Q000378:metabolism
D002712:Chlorides; D005290:Ferric Compounds; D009004:Monosaccharide Transport Proteins; D005947:Glucose; D005033:Ethylmaleimide; D001205:Ascorbic Acid; D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid; C024555:ferric chloride; D003683:Dehydroascorbic Acid