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Lipid Peroxidation Scavengers Prevent the Carbonylation of Cytoskeletal Brain Proteins Induced by Glutathione Depletion

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Summary

In this study, we investigated the possible link between lipid peroxidation (LPO) and the formation of protein carbonyls (PCOs) during depletion of brain glutathione (GSH). To this end, rat brain slices were incubated with the GSH depletor diethyl maleate (DEM) in the absence or presence of classical LPO scavengers: trolox, caffeic acid phenethyl ester (CAPE), and butylated hydroxytoluene (BHT). All three scavengers reduced DEM-induced lipid oxidation and protein carbonylation, suggesting that intermediates/products of the LPO pathway such as lipid hydroperoxides, 4-hydroxynonenal and/or malondialdehyde are involved in the process. Additional in vitro experiments revealed that, among these products, lipid hydroperoxides are most likely responsible for protein oxidation. Interestingly, BHT prevented the carbonylation of cytoskeletal proteins but not that of soluble proteins, suggesting the existence of different mechanisms of PCO formation during GSH depletion. In pull-down experiments, β-actin and α/β-tubulin were identified as major carbonylation targets during GSH depletion, although other cytoskeletal proteins such as neurofilament proteins and glial fibrillary acidic protein were also carbonylated. These findings may be important in the context of neurological disorders that exhibit decreased GSH levels and increased protein carbonylation such as Parkinson’s disease, Alzheimer’s disease, and multiple sclerosis.

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Abbreviations

AG:

Aminoguanidine

BHT:

Butylated hydroxytoluene

CAPE:

Caffeic acid phenethyl ester

CNS:

Central nervous system

DEM:

Diethyl maleate

ECL:

Enhanced chemioluminescence

GFAP:

Glial fibrillary acidic protein

GSH:

Glutathione

4-HNE:

4-hydoxy-2-nonenal

15-HPETE:

15-hydroperoxy-5,8,11,13-eicosanotetraenoic acid

Hy:

Hydralazine

LPO:

Lipid peroxidation

MDA:

Malondialdehyde

NFH:

Neurofilament heavy (200 kDa) protein

NFL:

Neurofilament light (69 kDa) protein

NFM:

Neurofilament medium (150 kDa) protein

PCO:

Protein carbonyl

RCO:

Reactive carbonyl

SDS-PAGE:

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis

TBARS:

Thiobarbituric acid reactive substances

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Acknowledgements

This work was supported by PHHS grants NS 47448 and IMSD GM 60201 from the National Institutes of Health.

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Authors and Affiliations

  1. Department of Cell Biology and Physiology, University of New Mexico—Health Sciences Center, Basic Medical Sciences Building, Room #151, 914 Camino de Salud, Albuquerque, NM, 87131-5218, USA

    Oscar A. Bizzozero, Savanna Reyes, Jennifer Ziegler & Suzanne Smerjac

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  1. Oscar A. Bizzozero
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  2. Savanna Reyes
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  3. Jennifer Ziegler
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  4. Suzanne Smerjac
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Correspondence to Oscar A. Bizzozero.

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Special issue in honor of Naren Banik.

Dr. Oscar Bizzozero dedicates this article to Dr. Naren Banik, outstanding scientist and colleague, in honor to his life-long achievements in the area of spinal cord injury and multiple sclerosis.

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Bizzozero, O.A., Reyes, S., Ziegler, J. et al. Lipid Peroxidation Scavengers Prevent the Carbonylation of Cytoskeletal Brain Proteins Induced by Glutathione Depletion. Neurochem Res 32, 2114–2122 (2007). https://doi.org/10.1007/s11064-007-9377-y

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