Increased Oxidation And Degradation Of Cytosolic Proteins In Alcohol-exposed Mouse Liver And Hepatoma Cells

Title

Increased Oxidation And Degradation Of Cytosolic Proteins In Alcohol-exposed Mouse Liver And Hepatoma Cells

Creator

Kim B J; Hood B L; Aragon R A; Hardwick Jr; Conrads T R; Veenstra T D; Song B J

Publisher

Proteomics

Date

2006
2006-02

Description

We recently developed a sensitive method using biotin-N-maleimide (biotin-NM) as a probe to positively identify oxidized mitochondrial proteins. In this study, biotin-NM was used to identify oxidized cytosolic proteins in alcohol-fed mouse livers. Alcohol treatment for 6 wk elevated the levels of CYP2E1 and nitrotyrosine, a marker of oxidative stress. Markedly increased levels of oxidized proteins were detected in alcohol-fed mouse livers compared to pair-fed controls. The biotin-NM-labeled oxidized proteins from alcohol-exposed mouse livers were subsequently purified with streptavidin-agarose and resolved on 2-DE. More than 90 silver-stained protein spots that displayed differential intensities on 2-D gels were identified by MS. Peptide sequence analysis revealed that many enzymes or proteins involved in stress response, chaperone activity, intermediary metabolism, and antioxidant defense systems such as peroxiredoxin were oxidized after alcohol treatment. Smaller fragments of many proteins were repeatedly detected only in alcohol-fed mice, indicating that many oxidized proteins after alcohol exposure were degraded. Immunoblot results showed that the level of oxidized peroxiredoxin (inactivated) was markedly increased in the alcohol-exposed mouse livers and ethanol-sensitive hepatoma cells compared to the corresponding controls. Our results may explain the underlying mechanism for cellular dysfunction and increased susceptibility to other toxic agents following alcohol-mediated oxidative stress.

Subject

alcoholism; apoptosis; Biochemistry & Molecular Biology; CYP2E1; cytochrome p-4502e1; Degradation; disease; ethanol; in-vitro; induced; injury; nitric-oxide; oxidative stress; oxidized proteins; peroxiredoxin; protein; protein oxidation; rat-liver; stress

Format

Journal Article or Conference Abstract Publication

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Rights

Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).

Pages

1250-1260

Issue

4

Volume

6

Citation

Kim B J; Hood B L; Aragon R A; Hardwick Jr; Conrads T R; Veenstra T D; Song B J, “Increased Oxidation And Degradation Of Cytosolic Proteins In Alcohol-exposed Mouse Liver And Hepatoma Cells,” NEOMED Bibliography Database, accessed June 21, 2021, https://neomed.omeka.net/items/show/10469.

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