Compartmental oxidation of thiol–disulphide redox couples during epidermal growth factor signalling

Halvey, Patrick J.; Watson, Walter H.; Hansen, Jason M.; Go, Young‐Mi; Samali, Afshin; Jones, Dean P.

doi:10.1042/bj20041829

Cited by 147 publications

(166 citation statements)

References 32 publications

Supporting

Mentioning

157

Contrasting

Order By: Relevance

“…With some oxidants (e.g. epidermal growth factor-induced ROS) (Halvey et al, 2005;Hansen et al, 2006a), Trx2 is not significantly oxidized. The reason(s) for the enhanced sensitivity of Trx2 to Cr(VI) are not known, but could indicate a more robust interaction of Cr(VI) with the mitochondrial thioredoxin system or a lesser ability of the mitochondrial system to restore or maintain Trx2 in its reduced state once it has been oxidized.…”

Section: Discussionmentioning

confidence: 99%

“…The redox states of Trx1 and Trx2 have been used to differentially assess the impacts of other oxidants on the thiol redox status of the cytosolic and mitochondrial compartments (Halvey et al, 2005;Hansen et al, 2006a). For the studies reported here, both insoluble and soluble Cr (VI) cause a dose-and time-dependent oxidation of Trx1 and Trx2 (Fig.…”

Section: Discussionmentioning

confidence: 99%

“…Adherent cells (60-70% confluent) were washed once in pre-warmed HBSS, and treated with Cr(VI) in HBSS at 37°C for the indicated times. Following treatment, they were washed once in HBSS and scraped into chilled 6 M guanidine-HCl, 100 mM Tris-HCl pH 8.3, 3 mM EDTA, 0.5 % Triton X-100, 50 mM iodoacetic acid (Halvey et al, 2005). The samples were briefly sonicated on ice to shear the DNA, and guanidine and unreacted iodoacetate were removed using Microspin G25 columns (GE Healthcare, Piscataway NJ).…”

Section: Trx1 Redox Blotsmentioning

confidence: 99%

“…The redox status of Trx1 was determined as adapted from Halvey et al (Halvey et al, 2005). Adherent cells (60-70% confluent) were washed once in pre-warmed HBSS, and treated with Cr(VI) in HBSS at 37°C for the indicated times.…”

Section: Trx1 Redox Blotsmentioning

confidence: 99%

“…Factors which enhance Trx oxidation would therefore be expected to interfere with Trx activity and could decrease cell survival. Various types of oxidants can result in Trx oxidation, including t-butyl hydroperoxide, ROS and the non-specific thiol oxidant diamide (Halvey et al, 2005;Chen et al, 2006;Hansen et al, 2006a). Arsenic and mercury can cause Trx oxidation (Hansen et al, 2006b) but the mechanisms involved are not clear.…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

Hexavalent chromium causes the oxidation of thioredoxin in human bronchial epithelial cells

2008

View full text Add to dashboard Cite

Hexavalent chromium [Cr(VI)] species such as chromates are cytotoxic. Inhalational exposure is a primary concern in many Cr-related industries and their immediate environments, and bronchial epithelial cells are directly exposed to inhaled Cr(VI). Chromates are readily taken up by cells and are reduced to reactive Cr species which may also result in the generation of reactive oxygen species (ROS). The thioredoxin (Trx) system has a key role in the maintenance of cellular thiol redox balance and is essential for cell survival. Cells normally maintain the cytosolic (Trx1) and mitochondrial (Trx2) thioredoxins largely in the reduced state. Redox western blots were used to assess the redox status of the thioredoxins in normal human bronchial epithelial cells (BEAS-2B) incubated with soluble Na 2 CrO 4 or insoluble ZnCrO 4 for different periods of time. Both chromates caused a doseand time-dependent oxidation of Trx2 and Trx1. Trx2 was more susceptible in that it could all be converted to the oxidized form, whereas a small amount of reduced Trx1 remained even after prolonged treatment with higher Cr concentrations. Only one of the dithiols, presumably the active site, of Trx1 was oxidized by Cr(VI). Cr(VI) did not cause significant GSH depletion or oxidation indicating that Trx oxidation does not result from a general oxidation of cellular thiols. With purified Trx and thioredoxin reductase (TrxR) in vitro, Cr(VI) also resulted in Trx oxidation. It was determined that purified TrxR has pronounced Cr(VI) reducing activity, so competition for electron flow from TrxR might impair its ability to reduce Trx. The in vitro data also suggested some direct redox interaction between Cr(VI) and Trx. The ability of Cr(VI) to cause Trx oxidation in cells could contribute to its cytotoxic effects, and could have important implications for cell survival, redoxsensitive cell signaling, and the cells' tolerance of other oxidant insults.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Trx1 Redox Blotsmentioning

confidence: 99%

Section: Trx1 Redox Blotsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Hexavalent chromium causes the oxidation of thioredoxin in human bronchial epithelial cells

2008

View full text Add to dashboard Cite

show abstract

Thioredoxin Redox Western Analysis

Jones

2009

CP Toxicology

View full text Add to dashboard Cite

Increasing evidence suggests that compartment-specific changes are important in redox signaling and control. Examination of thiol/disulfide redox changes in thioredoxin (Trx) family members including Trx1 in cytoplasm and nucleus and Trx2 in mitochondria should aid in the understanding of compartmentalized redox signaling mechanisms. Methods to quantify redox states of both Trx1 and Trx2 by redox western analysis and to further calculate redox potential using the Nernst equation are described in this unit. The procedures to measure redox states of Trx1 and Trx2 consist of three parts, derivatization, redox western blotting, and calculation of redox potentials. Derivatization of proteins with thiol-reactive reagents prior to redox western blotting prevents artifactual oxidation of protein thiols and allows separation of the reduced forms from oxidized forms of the protein. Consequently, the redox western analysis of Trx provides a convenient tool to estimate the redox state of different compartments and improve the understanding of redox signaling.

show abstract

Compartmentation of Redox Signaling and Control: Discrimination of Oxidative Stress in Mitochondria, Cytoplasm, Nuclei, and Endoplasmic Reticulum

Halvey

Hansen

Lash

et al. 2008

Drug‐Induced Mitochondrial Dysfunction

View full text Add to dashboard Cite

Compartmental oxidation of thiol–disulphide redox couples during epidermal growth factor signalling

Cited by 147 publications

References 32 publications

Hexavalent chromium causes the oxidation of thioredoxin in human bronchial epithelial cells

Hexavalent chromium causes the oxidation of thioredoxin in human bronchial epithelial cells

Thioredoxin Redox Western Analysis

Compartmentation of Redox Signaling and Control: Discrimination of Oxidative Stress in Mitochondria, Cytoplasm, Nuclei, and Endoplasmic Reticulum

Contact Info

Product

Resources

About