Glutathione redox potential in response to differentiation and enzyme inducers.
The reduced glutathione (GSH)/oxidized glutathione (GSSG) redox state is thought to function in signaling of detoxification gene expression, but also appears to be tightly regulated in cells under normal conditions. Thus it is not clear that the magnitude of change in response to physiologic stimuli is sufficient for a role in redox signaling under nontoxicologic conditions. The purpose of this study was to determine the change in 2GSH/GSSG redox during signaling of differentiation and increased detoxification enzyme activity in HT29 cells. We measured GSH, GSSG, cell volume, and cell pH, and we used the Nernst equation to determine the changes in redox potential Eh of the 2GSH/GSSG pool in response to the differentiating agent, sodium butyrate, and the detoxification enzyme inducer, benzyl isothiocyanate. Sodium butyrate caused a 60-mV oxidation (from -260 to -200 mV), an oxidation sufficient for a 100-fold change in protein dithiols:disulfide ratio. Benzyl isothiocyanate caused a 16-mV oxidation in control cells but a 40-mV oxidation (to -160 mV) in differentiated cells. Changes in GSH and mRNA for glutamate:cysteine ligase did not correlate with Eh; however, correlations were seen between Eh and glutathione S-transferase (GST) and nicotinamide adenine dinucleotide phosphate (NADPH):quinone reductase activities (N:QR). These results show that 2GSH/GSSG redox changes in response to physiologic stimuli such as differentiation and enzyme inducers are of a sufficient magnitude to control the activity of redox-sensitive proteins. This suggests that physiologic modulation of the 2GSH/GSSG redox poise could provide a fundamental parameter for the control of cell phenotype.
Kirlin
W G
WG
Department of Biochemistry, Emory University School of Medicine, Atlanta, GA 30322, USA.
Cai
J
J
Thompson
S A
SA
Diaz
D
D
Kavanagh
T J
TJ
Jones
D P
DP
D000230:Adenocarcinoma; D002087:Butyrates / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects*; D048429:Cell Size; D003110:Colonic Neoplasms; D004790:Enzyme Induction / Q000187:drug effects*; D015870:Gene Expression / Q000187:drug effects; D005721:Glutamate-Cysteine Ligase / Q000235:genetics; D005978:Glutathione / Q000378:metabolism*; D005982:Glutathione Transferase / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D017879:Isothiocyanates / Q000494:pharmacology; D016660:NAD(P)H Dehydrogenase (Quinone) / Q000378:metabolism; D010084:Oxidation-Reduction; D012333:RNA, Messenger / Q000378:metabolism; D014407:Tumor Cells, Cultured
D002087:Butyrates; D017879:Isothiocyanates; D012333:RNA, Messenger; C031403:benzyl isothiocyanate; D016660:NAD(P)H Dehydrogenase (Quinone); D005982:Glutathione Transferase; D005721:Glutamate-Cysteine Ligase; D005978:Glutathione