The mechanism of cysteine conjugate cytotoxicity in renal epithelial cells. Covalent binding leads to thiol depletion and lipid peroxidation

Qin Chen, T. W. Jones, P. C. Brown, J. L. Stevens

Research output: Contribution to journalArticle

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Abstract

Nephrotoxic cysteine conjugates kill cells after they are metabolized by the enzyme cysteine conjugate β-lyase to reactive fragments which bind to cellular macromolecules. We have investigated the cellular events which occur after the binding and lead ultimately to cell death in renal epithelial cells. Using S-(1,2-dichlorovinyl)-L-cysteine (DCVC) as a model conjugate, we found that the phenolic antioxidants N,N'-diphenyl-p-phenylenediamine (DPPD), butylated hydroxyanisole, butylated hydroxytoluene, propyl galate, and butylated hydroxyquinone, and the iron chelator deferoxamine inhibited the cytotoxicity significantly. Among the five antioxidants, DPPD was most potent. DPPD blocked DCVC toxicity over an extended time period, and the rescued cells remained functional as measured by protein synthetic activity. DPPD was able to block the toxicity of two other toxic cysteine conjugates S-(2-chloro-1,1,2-trifluoroethyl)-L-cysteine and S-(1,1,2,2-tetrafluoroethyl)-L-cysteine. In addition to LLC-PK1 cells, DPPD also protected freshly isolated rat kidney epithelial cells in suspension and in primary culture. In suspension cells, DPPD was effective at low doses of DCVC (25-50 μM) but not at high concentrations (250-500 μM). DPPD inhibition was not due to an inactivation of β-lyase or a decrease in the binding of [35S]DCVC metabolites to cellular macromolecules and occurred at a step after the activation of the toxins. During DCVC treatment, lipid peroxidation products were detectable prior to cell death. DPPD blocked lipid peroxidation over the whole time course. Depletion of nonprotein thiols also occurred prior to cell death. DPPD did not prevent the loss of nonprotein thiols. However, the sulfhydryl-reducing agent DTT blocked lipid peroxidation and toxicity at a step after the activation of DCVC. Therefore, it appears that cysteine conjugates kill renal epithelial cells by a combination of covalent binding, depletion of nonprotein thiols, and lipid peroxidation.

Original languageEnglish (US)
Pages (from-to)21603-21611
Number of pages9
JournalJournal of Biological Chemistry
Volume265
Issue number35
StatePublished - 1990
Externally publishedYes

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Cytotoxicity
Sulfhydryl Compounds
Lipid Peroxidation
Cysteine
Epithelial Cells
Kidney
Lipids
Cell death
Toxicity
Lyases
Cell Death
Macromolecules
Suspensions
Antioxidants
Chemical activation
LLC-PK1 Cells
Butylated Hydroxyanisole
Butylated Hydroxytoluene
Deferoxamine
4-phenylenediamine

ASJC Scopus subject areas

  • Biochemistry

Cite this

The mechanism of cysteine conjugate cytotoxicity in renal epithelial cells. Covalent binding leads to thiol depletion and lipid peroxidation. / Chen, Qin; Jones, T. W.; Brown, P. C.; Stevens, J. L.

In: Journal of Biological Chemistry, Vol. 265, No. 35, 1990, p. 21603-21611.

Research output: Contribution to journalArticle

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