Identification of Cysteine Residues Involved in Disulfide Formation in the Inactivation of Glutathione Transferase P-Form by Hydrogen Peroxide

1993 ◽  
Vol 300 (1) ◽  
pp. 137-141 ◽  
Author(s):  
H. Shen ◽  
S. Tsuchida ◽  
K. Tamai ◽  
K. Sato
2008 ◽  
Vol 19 (1) ◽  
pp. 226-236 ◽  
Author(s):  
Judith M. Müller ◽  
Dusanka Milenkovic ◽  
Bernard Guiard ◽  
Nikolaus Pfanner ◽  
Agnieszka Chacinska

The mitochondrial intermembrane space contains chaperone complexes that guide hydrophobic precursor proteins through this aqueous compartment. The chaperones consist of hetero-oligomeric complexes of small Tim proteins with conserved cysteine residues. The precursors of small Tim proteins are synthesized in the cytosol. Import of the precursors requires the essential intermembrane space proteins Mia40 and Erv1 that were proposed to form a relay for disulfide formation in the precursor proteins. However, experimental evidence for a role of Mia40 and Erv1 in the oxidation of intermembrane space precursors has been lacking. We have established a system to directly monitor the oxidation of precursors during import into mitochondria and dissected distinct steps of the import process. Reduced precursors bind to Mia40 during translocation into mitochondria. Both Mia40 and Erv1 are required for formation of oxidized monomers of the precursors that subsequently assemble into oligomeric complexes. Whereas the reduced precursors can diffuse back into the cytosol, the oxidized precursors are retained in the intermembrane space. Thus, oxidation driven by Mia40 and Erv1 determines vectorial transport of the precursors into the mitochondrial intermembrane space.


2016 ◽  
Vol 94 (4) ◽  
pp. 305-311 ◽  
Author(s):  
David J. Press ◽  
Thomas G. Back

A series of o-(hydroxymethyl)phenyl selenides containing single or multiple methoxy substituents was synthesized, and the rate at which each compound catalyzed the oxidation of benzyl thiol to its disulfide with excess hydrogen peroxide was measured. This assay provided the means for comparing the relative abilities of the selenides to mimic the antioxidant selenoenzyme glutathione peroxidase. The mechanism for catalytic activity involves oxidation of the selenides to their corresponding selenoxides with hydrogen peroxide, cyclization to spirodioxyselenuranes, followed by reduction with two equivalents of thiol to regenerate the original selenide with concomitant disulfide formation. A single p-methoxy group on each aryl moiety afforded the highest catalytic activity, while methoxy groups in the meta position had little effect compared to the unsubstituted selenide, and o-methoxy groups suppressed activity. The installation of multiple methoxy groups on each aryl moiety provided no improvement. These results can be rationalized on the basis of dominating mesomeric and steric effects of the p- and o-substituents, respectively.


2015 ◽  
Vol 396 (5) ◽  
pp. 465-482 ◽  
Author(s):  
Jan Riemer ◽  
Markus Schwarzländer ◽  
Marcus Conrad ◽  
Johannes M. Herrmann

Abstract Mitochondria are a major source of reactive oxygen species (ROS) in the cell, particularly of superoxide and hydrogen peroxide. A number of dedicated enzymes regulate the conversion and consumption of superoxide and hydrogen peroxide in the intermembrane space and the matrix of mitochondria. Nevertheless, hydrogen peroxide can also interact with many other mitochondrial enzymes, particularly those with reactive cysteine residues, modulating their reactivity in accordance with changes in redox conditions. In this review we will describe the general redox systems in mitochondria of animals, fungi and plants and discuss potential target proteins that were proposed to contain regulatory thiol switches.


2014 ◽  
Vol 42 (1) ◽  
pp. 42-46 ◽  
Author(s):  
Rachel E. Martin ◽  
Zhenbo Cao ◽  
Neil J. Bulleid

Hydrogen peroxide (H2O2) can act as a signalling molecule affecting the cell cycle as well as contributing towards the oxidative stress response. The primary target of this molecule is oxidation-sensitive cysteine residues in proteins such as protein tyrosine phosphatases. The cell has robust mechanisms to remove H2O2 that need to be regulated for H2O2 to react with and modify protein thiols. In particular, the family of peroxiredoxins are capable of the rapid removal of even trace amounts of this molecule. It has been suggested that the inactivation of peroxiredoxins by hyperoxidation may allow H2O2 levels to increase in cells and thereby modify critical thiol groups in proteins. We have been studying how the H2O2 produced during disulfide formation in the ER (endoplasmic reticulum) is metabolized and have shown that ER-resident peroxiredoxin IV not only can remove H2O2, but also contributes to de novo disulfide formation. In the present article, we review recent data on the structure and function of this enzyme as well as its sensitivity to hyperoxidation.


2015 ◽  
Vol 396 (5) ◽  
pp. 389-399 ◽  
Author(s):  
Lars I. Leichert ◽  
Tobias P. Dick

Abstract A few small-molecule oxidants, most notably hydrogen peroxide, can act as messengers in signal transduction. They trigger so-called ‘thiol switches’, cysteine residues that are reversibly oxidized to transiently change the functional properties of their host proteins. The proteome-wide identification of functionally relevant ‘thiol switches’ is of significant interest. Unfortunately, prediction of redox-active cysteine residues on the basis of surface accessibility and other computational parameters appears to be of limited use. Proteomic thiol labeling approaches remain the most reliable strategy to discover new thiol switches in a hypothesis-free manner. We discuss if and how genomic knock-in strategies can help establish the physiological relevance of a ‘thiol switch’ on the organismal level. We conclude that surprisingly few attempts have been made to thoroughly verify the physiological relevance of thiol-based redox switches in mammalian model organisms.


2020 ◽  
Vol 74 ◽  
pp. 1-11
Author(s):  
Marzena Gutowicz

Since the brain contains a large amount of polyunsaturated fatty acids, consumes up to 20% of oxygen used by the whole body and exhibits low antioxidants activity, it seems to be especially vulnerable to oxidative stress. The most important antioxidant enzymes are superoxide dismutase (SOD), which catalyze the dismutation of superoxide anion to hydrogen peroxide, catalase (CAT), which converts toxic hydrogen peroxide to water and oxygen, and glutathione peroxidase (Se-GSHPx), which reduces hydrogen peroxide and organic peroxides with glutathione as the cofactor. Among other detoxifying enzymes, the most significant is glutathione transferase (GST), which shows detoksyvarious catalytic activities allowing for removal of xenobiotics, reducing organic peroxides and oxidized cell components. One of the most important brain nonenzymatic antioxidants is reduced glutathione (GSH), which (individually or in cooperation with peroxidases) participates in the reduction of free radicals, repair of oxidative damage and the regeneration of other antioxidants, such as ascorbate or tocopherol. Glutathione as a cosubstrate of glutathione transferase scavenges toxic electrophilic compounds. Although the etiology of the major neurodegenerative diseases are unknown, numerous data suggest that reactive oxygen species play an important role. Even a small change in the level of antioxidants can leads to the many disorders in the CNS.


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