Plasma protein thiol oxidation and carbonyl formation in chronic renal failure.
BACKGROUND
Myeloperoxidase-catalyzed oxidative pathways have recently been identified as an important cause of oxidant stress in uremia and hemodialysis (HD), and can lead to plasma protein oxidation. We have examined patterns of plasma protein oxidation in vitro in response to hydrogen peroxide (H2O2) and hypochlorous acid (HOCl). We measured thiol oxidation, amine oxidation, and carbonyl concentrations in patients on chronic maintenance HD compared with patients with chronic renal failure (CRF) and normal volunteers. We have also examined the effect of the dialysis procedure on plasma protein oxidation using biocompatible and bioincompatible membranes.


METHODS
Plasma proteins were assayed for the level of free thiol groups using spectrophotometry, protein-associated carbonyl groups by enzyme-linked immunosorbent assay, and oxidation of free amine groups using a fluorescent spectrophotometer.


RESULTS
In vitro experiments demonstrate HOCl oxidation of thiol groups and increased carbonyl formation. In vivo, there are significant differences in plasma-free thiol groups between normal volunteers (279 +/- 12 micromol/L), CRF patients (202 +/- 20 micromol/L, P = 0.005) and HD patients (178 +/- 18 micromol/L, P = 0.0001). There are also significant differences in plasma protein carbonyl groups between normal volunteers (0.76 +/- 0.51 micromol/L), CRF patients (13.73 +/- 4.45 micromol/L, P = 0.015), and HD patients (16.95 +/- 2.62 micromol/L, P = 0.0001). There are no significant differences in amine group oxidation. HD with both biocompatible and bioincompatible membranes restored plasma protein thiol groups to normal levels, while minimally affecting plasma protein carbonyl expression.


CONCLUSIONS
First, both CRF and HD patients have increased plasma protein oxidation manifested by oxidation of thiol groups and formation of carbonyl groups. Second, HD with biocompatible and bioincompatible membranes restored plasma protein thiol groups to normal levels. Third, these experiments suggest that there is a dialyzable low molecular weight toxin found in uremia that is responsible for plasma protein oxidation.
Himmelfarb
J
J
Maine Medical Center, Portland, and Maine Medical Center Research Institute, South Portland, Maine 04102, USA. himmej@mail.mmc.org
McMonagle
E
E
McMenamin
E
E
D000588:Amines / Q000378:metabolism; D001672:Biocompatible Materials; D001798:Blood Proteins / Q000378:metabolism*; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D006997:Hypochlorous Acid / Q000378:metabolism; D066298:In Vitro Techniques; D007676:Kidney Failure, Chronic / Q000378:metabolism* / Q000628:therapy; D008567:Membranes, Artificial; D016877:Oxidants / Q000494:pharmacology; D010084:Oxidation-Reduction; D018384:Oxidative Stress / Q000187:drug effects / Q000502:physiology; D010586:Phagocytes / Q000378:metabolism; D006435:Renal Dialysis; D013438:Sulfhydryl Compounds / Q000378:metabolism*; D014511:Uremia / Q000378:metabolism / Q000628:therapy
D000588:Amines; D001672:Biocompatible Materials; D001798:Blood Proteins; D008567:Membranes, Artificial; D016877:Oxidants; D013438:Sulfhydryl Compounds; D006997:Hypochlorous Acid; D006861:Hydrogen Peroxide