Stimulation of a human erythrocyte membrane ATPase by glutathione conjugates.
An ATP-dependent transport process for S-(2,4-dinitrophenyl) glutathione (Dnp-SG) mediated by a novel ATPase designated as Dnp-SG ATPase has been demonstrated in human erythrocytes (LaBelle et al., FEBS Lett. 228, 53-51, 1988). In order to investigate whether the Dnp-SG ATPase system represents a generalized mechanism for the transport of xenobiotic conjugates of glutathione (GSH), stimulation of this ATPase by different GSH conjugates was studied in membrane vesicles prepared from human erythrocytes. Kinetic parameters for several GSH conjugates including S-(methyl)glutathione, S-(n-propyl)glutathione, S-(n-pentyl)glutathione, S-(n-decyl)glutathione, S-(p-chlorophenacyl)glutathione, S-(p-nitrobenzyl)glutathione, and the GSH conjugate of 9,10-epoxystearic acid were determined in order to evaluate their affinity for Dnp-SG ATPase. These studies reveal that all these conjugates stimulated Dnp-SG ATPase of human erythrocyte membrane. The apparent Km values of Dnp-SG ATPase for different conjugates were found to be in the range of 0.26-0.66 mM with Vmax values ranging from 0.55 to 4.44 nmol/min/mg protein. The results of these studies indicate that erythrocyte membrane Dnp-SG ATPase represents a generalized mechanism for the transport of GSH conjugates formed with xenobiotics as well as with the endogenously generated electrophilic compounds such as epoxystearic acid. It is suggested that Dnp-SG ATPase in conjunction with GSH and GSH S-transferase may play an important role in the protection of erythrocytes from exogenous as well as endogenous electrophilic toxicants.
Sharma
R
R
Department of Human Biological Chemistry and Genetics, University of Texas Medical Branch, Galveston 77550.
Gupta
S
S
Ahmad
H
H
Ansari
G A
GA
Awasthi
Y C
YC
D000251:Adenosine Triphosphatases / Q000096:biosynthesis*; D000255:Adenosine Triphosphate / Q000493:pharmacokinetics; D001693:Biological Transport, Active; D002462:Cell Membrane / Q000187:drug effects*; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004912:Erythrocytes / Q000201:enzymology / Q000648:ultrastructure; D005978:Glutathione / Q000031:analogs & derivatives* / Q000493:pharmacokinetics; D006801:Humans; D066298:In Vitro Techniques
C030190:S-(2,4-dinitrophenyl)glutathione; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases; D005978:Glutathione