Gulonolactone oxidase activity-dependent intravesicular glutathione oxidation in rat liver microsomes.
The orientation of gulonolactone oxidase activity was investigated in rat liver microsomes. Ascorbate formation upon gulonolactone addition resulted in higher intravesicular than extravesicular ascorbate concentrations in native microsomal vesicles. The intraluminal ascorbate accumulation could be prevented or the accumulated ascorbate could be released by permeabilising the vesicles with the pore-forming alamethicin. The formation of the other product of the enzyme, hydrogen peroxide caused the preferential oxidation of intraluminal glutathione in glutathione-loaded microsomes. In conclusion, these results suggest that the orientation of the active site of gulonolactone oxidase is intraluminal and/or the enzyme releases its products towards the lumen of the endoplasmic reticulum.
Department of Medical Chemistry, Molecular Biology and Pathobiochemistry, Semmelweis University of Medicine, Budapest, Hungary.
F
F
Puskás
L
L
Braun
M
M
Csala
T
T
Kardon
P
P
Marcolongo
A
A
Benedetti
J
J
Mandl
G
G
Bánhegyi
D000408:Alamethicin / Q000494:pharmacology; D000818:Animals; D001205:Ascorbic Acid / Q000378:metabolism; D004789:Enzyme Activation; D005978:Glutathione / Q000378:metabolism*; D019803:Glutathione Disulfide / Q000378:metabolism; D050544:L-Gulonolactone Oxidase; D008027:Light; D008297:Male; D008862:Microsomes, Liver / Q000201:enzymology*; D010084:Oxidation-Reduction; D051381:Rats; D017207:Rats, Sprague-Dawley; D012542:Scattering, Radiation; D013400:Sugar Acids / Q000378:metabolism; D013401:Sugar Alcohol Dehydrogenases / Q000378:metabolism*; D014475:Uncoupling Agents / Q000494:pharmacology
D013400:Sugar Acids; D014475:Uncoupling Agents; D000408:Alamethicin; C007895:gulonolactone; D013401:Sugar Alcohol Dehydrogenases; D050544:L-Gulonolactone Oxidase; D005978:Glutathione; D001205:Ascorbic Acid; D019803:Glutathione Disulfide