The effect of H2O2 upon thioredoxin-enriched lens epithelial cells.
Thioredoxin, a dithiol polypeptide, has been examined as a potential contributor to the recovery of lens epithelial cells from oxidative insult. It is reported that Escherichia coli thioredoxin can (a) effectively reduce lens-soluble protein disulfide bonds generated by H2O2, (b) restore to its initial activity H2O2-inactivated glyceraldehyde-3-phosphate dehydrogenase, (c) act as an effective source of reducing potential for lens methionine sulfoxide peptide reductase, and (d) act as a free radical quencher based on studies with a stable free radical system generated by ascorbic acid and 2,6-dimethoxy-p-benzoquinone. Thioredoxin is much more effective than dithiothreitol in restoring glyceraldehyde-3-phosphate dehydrogenase activity and as a cofactor for methionine sulfoxide peptide reductase. Upon incubation with epithelial cells, thioredoxin can be observed in the cell using rocket immunoelectrophoresis. These cells recover from H2O2 insult more rapidly than control cell preparations based upon 1) analyses of plasma membrane-related activities: leucine and 86Rb uptake and 2) analyses of parameters primarily related to the internal cell metabolism: ATP concentration and glyceraldehyde-3-phosphate dehydrogenase activity. Analysis of thioredoxin in cell preparations indicates that only about 9% is in the reduced state implying a low effective concentration of the polypeptide. The experiments suggest that low levels of thioredoxin may significantly increase the ability of lens epithelial cells to recover from exposure to H2O2.
Spector
A
A
Department of Ophthalmology, College of Physicians and Surgeons, Columbia University, New York, New York 10032.
Yan
G Z
GZ
Huang
R R
RR
McDermott
M J
MJ
Gascoyne
P R
PR
Pigiet
V
V
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism*; D001693:Biological Transport, Active / Q000187:drug effects; D002417:Cattle; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D004578:Electron Spin Resonance Spectroscopy; D004848:Epithelium / Q000187:drug effects / Q000378:metabolism; D004926:Escherichia coli / Q000187:drug effects / Q000378:metabolism; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D007700:Kinetics; D007908:Lens, Crystalline / Q000187:drug effects / Q000378:metabolism*; D007930:Leucine / Q000378:metabolism; D010084:Oxidation-Reduction; D012413:Rubidium / Q000378:metabolism; D013879:Thioredoxins / Q000378:metabolism* / Q000494:pharmacology
D001426:Bacterial Proteins; D013879:Thioredoxins; D000255:Adenosine Triphosphate; D006861:Hydrogen Peroxide; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D007930:Leucine; D012413:Rubidium