Transport of bimane-S-glutathione in human erythrocytes.
Export of glutathione S-conjugate of bimane (BSG) was studied in human erythrocytes. Characteristics of the BSG transport is similar to that of dinitrophenyl-S-glutathione (DNP-SG). BSG transport has two kinetic components, one of high affinity and low capacity (Km = 7.4 +/- 0.2 mumol/ml cells, Vm = 2.7 +/- 0.1 nmol/min per ml RBC) and another of low affinity and high capacity (Km = 242 +/- 8 mumol/ml cells, Vm = 9.6 +/- 1.6 nmol/min per ml RBC). BSG export is inhibited by vanadate (Ki = 65 +/- 6 microM) and fluoride (Ki = 11.4 +/- 0.8 mM). Activation energy of the transport is 67 +/- 7 kJ/mol. BSG transport is independent of membrane potential; its rate increases with pH in the pH range of 6-8, in line with the assumption that the anionic conjugate is cotransported with proton. BSG import to erythrocyte membrane inside-out vesicles is stimulated by ATP. Fluorimetric measurements of BSG export require low amounts of cells and may also be useful for other cell types as an alternative to studies of glutatione S-conjugate transport using radioactive substrates.
Pułaski
L
L
Department of Molecular Biophysics, University of Lódź, Poland.
Bartosz
G
G
D001692:Biological Transport / Q000187:drug effects; D001643:Bridged Bicyclo Compounds / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D004912:Erythrocytes / Q000378:metabolism*; D005033:Ethylmaleimide / Q000494:pharmacology; D005459:Fluorides / Q000494:pharmacology; D005978:Glutathione / Q000031:analogs & derivatives* / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D013696:Temperature; D014638:Vanadates / Q000494:pharmacology
D001643:Bridged Bicyclo Compounds; C080120:glutathione-bimane; C030190:S-(2,4-dinitrophenyl)glutathione; D014638:Vanadates; D005978:Glutathione; D005033:Ethylmaleimide; D005459:Fluorides