The not so identical twins: (dis)similarities between reactive electrophile and oxidant sensing and signaling

Long, Marcus J. C.; Huang, Kuan‐Ting; Aye, Yimon

doi:10.1039/d1cs00467k

Cited by 7 publications

(8 citation statements)

References 75 publications

(75 reference statements)

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“…Identifying molecular mechanisms responsible for modulation of cellular phenotype by endogenous metabolites is critical for understanding the mechanisms by which metabolism contributes to overall health and disease. 89 Indeed, DNA−protein cross-links induced by endogenous compounds represent an important class of phenotype-altering event and many of the proteins participating in these cross-links remain unidentified due to challenges in DPC isolation and characterization. In this study, we investigated the formation of DNA−protein crosslinks upon exposure of human cells in culture to electrophilic cellular metabolite MGO.…”

Section: ■ Discussionmentioning

confidence: 99%

Endogenous Cellular Metabolite Methylglyoxal Induces DNA–Protein Cross-Links in Living Cells

Hurben,

Zhang,

Galligan

et al. 2024

ACS Chem. Biol.

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Methylglyoxal (MGO) is an electrophilic α-oxoaldehyde generated endogenously through metabolism of carbohydrates and exogenously due to autoxidation of sugars, degradation of lipids, and fermentation during food and drink processing. MGO can react with nucleophilic sites within proteins and DNA to form covalent adducts. MGO-induced advanced glycation end-products such as protein and DNA adducts are thought to be involved in oxidative stress, inflammation, diabetes, cancer, renal failure, and neurodegenerative diseases. Additionally, MGO has been hypothesized to form toxic DNA−protein crosslinks (DPC), but the identities of proteins participating in such cross-linking in cells have not been determined. In the present work, we quantified DPC formation in human cells exposed to MGO and identified proteins trapped on DNA upon MGO exposure using mass spectrometry-based proteomics. A total of 265 proteins were found to participate in MGO-derived DPC formation including gene products engaged in telomere organization, nucleosome assembly, and gene expression. In vitro experiments confirmed DPC formation between DNA and glyceraldehyde-3-phosphate dehydrogenase (GAPDH), as well as histone proteins H3.1 and H4. Collectively, our study provides the first evidence for MGO-mediated DNA−protein cross-linking in living cells, prompting future studies regarding the relevance of these toxic lesions in cancer, diabetes, and other diseases linked to elevated MGO levels.

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Section: ■ Discussionmentioning

confidence: 99%

Endogenous Cellular Metabolite Methylglyoxal Induces DNA–Protein Cross-Links in Living Cells

Hurben,

Zhang,

Galligan

et al. 2024

ACS Chem. Biol.

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Section: Control Scenarios For Pseudo-intramolecular Electrophile Del...mentioning

confidence: 99%

“…Lipid-derived electrophiles interact with target proteins through cysteine-specific labeling, at least in many of the instances studied, especially when under kinetic control. 33 In many of these cases, it transpires that the targeted cysteine is dispensable for activity/function of the target protein. 19 Thus, performing delivery to a Halo−POI mutant in which the acceptor cysteine has been mutated to serine or alanine leads to no labeling of the POI, and the LDE is released to the bulk of the cells (Figure 4).…”

Section: Pseudo-intramolecular Electrophile Delivery Processesmentioning

confidence: 99%

Hiding in Plain Sight: The Issue of Hidden Variables

2022

Self Cite

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Here we discuss “hidden variables”, which are typically introduced during an experiment as a consequence of the application of two independent variables together to create a stimulus. With increased sophistication in modern chemical biology tools and related precision interrogation techniques, hidden variables have become integral to many chemical biologists’ routine experiments. For instance, they can appear in the use of light-activatable chemical probes (e.g., μMap, T-REX), or stimulus-induced enzyme activation (e.g., APEX). Unfortunately, control experiments assess only how independent variables affect measured outcomes and not the multiple differences between the two independent variables and the twain. We outline ways to account for potential hidden variables in experimental design and data interpretation as a means to aid developers of new methods, particularly those involving light-driven techniques, chemical activation, or biorthogonal chemistries, to better incorporate well-controlled procedures.

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“…For instance, spatial arrangements and sterics may enhance some cross-linking reactions and prevent others from occurring, even when disulfide formation is still possible. Indeed, while there is often overlap between oxidizable cysteines and those frequently targeted by electrophiles, protein thiols targeted by different classes of reactive species do vary in some cases . Therefore, follow-up experiments to confirm direct mixed disulfide formation may be warranted.…”

Section: Strategies For Identifying Proteins Involved In Redox Relaysmentioning

confidence: 99%

“…Indeed, while there is often overlap between oxidizable cysteines and those frequently targeted by electrophiles, 175 species do vary in some cases. 188 Therefore, follow-up experiments to confirm direct mixed disulfide formation may be warranted.…”

Section: Kinetic Trapping Approaches Using Mutantmentioning

confidence: 99%

Experimental Approaches for Investigating Disulfide-Based Redox Relays in Cells

West

2022

Chem. Res. Toxicol.

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Reversible oxidation of cysteine residues within proteins occurs naturally during normal cellular homeostasis and can increase during oxidative stress. Cysteine oxidation often leads to the formation of disulfide bonds, which can impact protein folding, stability, and function. Work in both prokaryotic and eukaryotic models over the past five decades has revealed several multiprotein systems that use thiol-dependent oxidoreductases to mediate disulfide bond reduction, formation, and/or rearrangement. Here, I provide an overview of how these systems operate to carry out disulfide exchange reactions in different cellular compartments, with a focus on their roles in maintaining redox homeostasis, transducing redox signals, and facilitating protein folding. Additionally, I review thiol-independent and thiol-dependent approaches for interrogating what proteins partner together in such disulfide-based redox relays. While the thiol-independent approaches rely either on predictive measures or standard procedures for monitoring protein−protein interactions, the thiol-dependent approaches include direct disulfide trapping methods as well as thiol-dependent chemical cross-linking. These strategies may prove useful in the systematic characterization of known and newly discovered disulfide relay mechanisms and redox switches involved in oxidant defense, protein folding, and cell signaling.

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The not so identical twins: (dis)similarities between reactive electrophile and oxidant sensing and signaling

Abstract: In this tutorial review, we compare and contrast the chemical mechanisms of electrophile/oxidant sensing, and the molecular mechanisms of signal propagation.

Cited by 7 publications

References 75 publications

Endogenous Cellular Metabolite Methylglyoxal Induces DNA–Protein Cross-Links in Living Cells

Endogenous Cellular Metabolite Methylglyoxal Induces DNA–Protein Cross-Links in Living Cells

Hiding in Plain Sight: The Issue of Hidden Variables

Experimental Approaches for Investigating Disulfide-Based Redox Relays in Cells

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