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Biochim Biophys Acta. 2014 Feb;1840(2):913-22. doi: 10.1016/j.bbagen.2013.09.003. Epub 2013 Sep 8.

Detection of electrophile-sensitive proteins.

Author information

1
Department of Pathology, Center for Free Radical Biology, University of Alabama at Birmingham, Birmingham, AL, USA.

Abstract

BACKGROUND:

Redox signaling is an important emerging mechanism of cellular function. Dysfunctional redox signaling is increasingly implicated in numerous pathologies, including atherosclerosis, diabetes, and cancer. The molecular messengers in this type of signaling are reactive species which can mediate the post-translational modification of specific groups of proteins, thereby effecting functional changes in the modified proteins. Electrophilic compounds comprise one class of reactive species which can participate in redox signaling. Electrophiles modulate cell function via formation of covalent adducts with proteins, particularly cysteine residues.

SCOPE OF REVIEW:

This review will discuss the commonly used methods of detection for electrophile-sensitive proteins, and will highlight the importance of identifying these proteins for studying redox signaling and developing novel therapeutics.

MAJOR CONCLUSIONS:

There are several methods which can be used to detect electrophile-sensitive proteins. These include the use of tagged model electrophiles, as well as derivatization of endogenous electrophile-protein adducts.

GENERAL SIGNIFICANCE:

In order to understand the mechanisms by which electrophiles mediate redox signaling, it is necessary to identify electrophile-sensitive proteins and quantitatively assess adduct formation. Strengths and limitations of these methods will be discussed. This article is part of a Special Issue entitled Current methods to study reactive oxygen species - pros and cons and biophysics of membrane proteins. Guest Editor: Christine Winterbourn.

KEYWORDS:

15-deoxy-Δ(12,14)-prostaglandin J(2); 15d-PGJ(2); 4-HNE; 4-hydroxynonenal; BD-15d-PGJ(2); BODIPY FL-15-deoxy-Δ(12,14)-prostaglandin J(2); DNA; DNPH; ELISA; Electrophile; Ep; GSH; GST; IAM; Keap1; Kelch-like enoyl-CoA hydratase-associated protein 1; N-acetylcysteine; N-ethylmaleimide; NAC; NEM; Nrf2; Oxidative post-translational modification; Proteomics; Reactive species; Redox signaling; Thiol; biotin 15-deoxy-Δ(12,14)-prostaglandin J(2); bt-15d-PGJ(2); deoxyribonucleic acid; dinitrophenylhydrazine; electrophile; enzyme-linked immunosorbent assay; glutathione; glutathione S-transferase; iodoacetamide; mito-15d-PGJ(2); mitochondrially targeted 15-deoxy-Δ(12,14)-prostaglandin J(2); nuclear factor-erythroid 2-related factor 2

PMID:
24021887
PMCID:
PMC3858512
DOI:
10.1016/j.bbagen.2013.09.003
[Indexed for MEDLINE]
Free PMC Article

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