Renal cysteine conjugate β-lyase-mediated toxicity studied with primary cultures of human proximal tubular cells

Chen, John C.; Stevens, James; Trifillis, Anna L.; Jones, Thomas W.

doi:10.1016/0041-008x(90)90319-p

Cited by 55 publications

(38 citation statements)

References 36 publications

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“…Several characteristics of DCVC make this an ideal model compound to study molecular mechanisms of nephrotoxicity. DCVC is specifically taken up by renal proximal tubular cells both in vitro and in vivo where it is bio-activated by the renal cysteine conjugate ␤-lyase activity, resulting in the formation of reactive intermediates that cause cell injury (23,25). PTC injury is associated with loss of focal adhesions and cell adhesion, which is followed by caspase activation and the onset of apoptosis (20,26,27).…”

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confidence: 99%

Heat Shock Protein 27 Is the Major Differentially Phosphorylated Protein Involved in Renal Epithelial Cellular Stress Response and Controls Focal Adhesion Organization and Apoptosis

Graauw

Tijdens

Cramer

et al. 2005

Journal of Biological Chemistry

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We used two-dimensional difference gel electrophoresis to determine early changes in the stress-response pathways that precede focal adhesion disorganization linked to the onset of apoptosis of renal epithelial cells. Treatment of LLC-PK1 cells with the model nephrotoxicant 1,2-(dichlorovinyl)-L-cysteine (DCVC) resulted in a >1.5-fold up-and down-regulation of 14 and 9 proteins, respectively, preceding the onset of apoptosis. Proteins included those involved in metabolism, i.e. aconitase and pyruvate dehydrogenase, and those related to stress responses and cytoskeletal reorganization, i.e. cofilin, Hsp27, and ␣-b-crystallin. The most prominent changes were found for Hsp27, which was related to a pI shift in association with an altered phosphorylation status of serine residue 82. Although both p38 and JNK were activated by DCVC, only inhibition of p38 with SB203580 reduced Hsp27 phosphorylation, which was associated with accelerated reorganization of focal adhesions, cell detachment, and apoptosis. In contrast, inhibition of JNK with SP600125 maintained cell adhesion as well as protection against apoptosis. Active JNK co-localized at focal adhesions after DCVC treatment in a FAK-dependent manner. Inhibition of active JNK localization at focal adhesions did not prevent DCVC-induced phosphorylation of Hsp27. Overexpression of a phosphorylationdefective mutant Hsp27 acted as a dominant negative and accelerated the DCVC-induced changes in the focal adhesions as well as the onset of apoptosis. Our data fit a model whereby early p38 activation results in a rapid phosphorylation of Hsp27, a requirement for proper maintenance of cell adhesion, thus suppressing renal epithelial cell apoptosis.

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confidence: 99%

Heat Shock Protein 27 Is the Major Differentially Phosphorylated Protein Involved in Renal Epithelial Cellular Stress Response and Controls Focal Adhesion Organization and Apoptosis

Graauw

Tijdens

Cramer

et al. 2005

Journal of Biological Chemistry

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“…Glutathione conjugates typically undergo enterohepatic circulation and can be ultimately cleaved to their cysteine S-conjugates, which can enter renal cells to produce toxicity (Anders et al, 1988;Koob and Dekant, 1991). Since the cysteine S-conjugates DCVC and CTFC are toxic to primary cultures of human RPT cells (Chen et al, 1990), the pathways responsible for their entry in the human RPT could play a critical role in the development of cell injury. The transport pathways involved in the uptake of cysteine conjugates by human kidney are not established, however.…”

Section: Discussionmentioning

confidence: 99%

“…Although many cysteine S-conjugates appear to be nephrotoxic and even nephrocarcinogenic in humans (Chen et al, 1990;Birner et al, 1993Birner et al, , 1997Bruning and Bolt, 2000;Lash et al, 2000), the role of the human organic transporter 1 (hOAT1) in the transport of these substrates has been not shown. Although the negatively charged N-acetyl derivatives of DCVC (NAC-DCVC) and of CTFC (NAC-CTFC) have been shown to be substrates for rat Oat1 (Pombrio et al, 2001), the interaction of the parent zwitterionic cysteine conjugate DCVC or CTFC with OAT1 is not known.…”

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confidence: 99%

Interaction of Cysteine Conjugates with Human and Rabbit Organic Anion Transporter 1

Groves

Munoz

Bahn

et al. 2003

J Pharmacol Exp Ther

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Organic anion (OA) transport mediates accumulation of the zwitterionic nephrotoxic cysteine S-conjugates S-dichlorovinylcysteine (DCVC) and S-chlorotrifluoroethylcysteine (CTFC) in the rabbit renal proximal tubule (RPT). Although these cysteine conjugates are nephrotoxic to the human RPT, neither the role of OA transport nor the specific OA transport pathway(s) involved in cysteine conjugate accumulation are known. Since the OAT1 transporter has the characteristics of para-aminokippurate (PAH) transport that closely correlate to the native RPT, we examined the interaction of DCVC, CTFC, and the nontoxic benzothiazolylcysteine (BTC) with PAH transport mediated by human OAT1 and rabbit Oat1 expressed in Chinese hamster ovary and COS7 heterologous expression systems, respectively. Although the K m values for PAH uptake by hOAT1 and rbOat1 (8.9 Ϯ 3.6 and 20.7 Ϯ 8 M, respectively) were 5-to 10-fold less than the K m for peritubular PAH transport into rabbit RPT, the IC 50 values for DCVC, CTFC, and BTC inhibition of PAH uptake mediated by either hOAT1 or rbOat1 were similar between these two transporters and to the IC 50 values for these conjugates measured in rabbit RPT. The IC 50 for inhibition of hOAT1-and rbOat1-mediated PAH uptake by the hydrophobic conjugate BTC was more than 5-fold lower than the IC 50 values seen with DCVC and CTFC, suggesting that hydrophobicity increases the affinity of OAT1 for cysteine conjugates. Finally, preloading cells transfected with hOAT1 with BTC significantly trans-stimulated the uptake of PAH, consistent with the conclusion that BTC and, hence, other cysteine S-conjugates are substrates for hOAT1.

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“…In view of the central role of VDAC2 as an inhibitor of the activation of the proapoptotic protein BAK, and the mitochondrial apoptotic pathway, their data point to a hitherto unrecognized mechanism by which TCDD might affect cellular homeostasis and survival. Another example is 1,2-dichlorovinyl-L-cysteine (DCVC), which belongs to the group of nephrotoxic halogenated alkene S-cysteine conjugates and is specifically taken up by renal proximal tubular cells both in vitro and in vivo where it is bio-activated by the renal cysteine conjugate-lyase activity, resulting in the formation of reactive intermediates that cause cell injury [42]. De Graauw et al [43] used 2D-DIGE to determine early changes in the onset of apoptosis of renal epithelial cells induced by DCVC.…”

Section: Chemical Compound-induced Apoptosismentioning

confidence: 99%

Current advances in the application of proteomics in apoptosis research

Wang

Chen

2011

Sci. China Life Sci.

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Apoptosis, or programmed cell death, is a complex, genetically-determined process involved in the development and maintenance of homeostasis in multicellular organisms. Dysregulation of apoptosis has been implicated in a number of diseases, including cancer and autoimmune disease. Thus, the investigation of apoptotic regulation has evoked considerable interest. Many apoptotic proteins have been shown to be post-translationally modulated, such as by protein cleavage, translocation, protein-protein interaction, and various post-translational modifications, which fall precisely within the range of proteomic analysis. Recently, contemporary proteomic technologies have achieved significant advances and have accelerated research in functional and chemical proteomics, which have been applied to the field of apoptosis research and have the potential to be a driving force for the field. This review highlights some of the major achievements in the application of proteomics in apoptosis research and discusses new directions and challenges for the near future.

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Renal cysteine conjugate β-lyase-mediated toxicity studied with primary cultures of human proximal tubular cells

Cited by 55 publications

References 36 publications

Heat Shock Protein 27 Is the Major Differentially Phosphorylated Protein Involved in Renal Epithelial Cellular Stress Response and Controls Focal Adhesion Organization and Apoptosis

Heat Shock Protein 27 Is the Major Differentially Phosphorylated Protein Involved in Renal Epithelial Cellular Stress Response and Controls Focal Adhesion Organization and Apoptosis

Interaction of Cysteine Conjugates with Human and Rabbit Organic Anion Transporter 1

Current advances in the application of proteomics in apoptosis research

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