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Updated: May 30, 2026
Synthesizing Amino Acids Modified with Reactive Carbonyls in Silico to Assess Structural Effects Using Molecular Dynamics Simulations
Published on: April 26, 2024
Protein targets for carbonylation by 4-hydroxy-2-nonenal in rat liver mitochondria
Jia Guo1, Katalin Prokai-Tatrai, Vien Nguyen
1Department of Molecular Biology and Immunology,University of North Texas Health Science Center, 3500 Camp Bowie Boulevard, Fort Worth, TX 76101, USA.
Abstract:
Protein carbonylation has been associated with various pathophysiological processes. A representative reactive carbonyl species (RCS), 4-hydroxy-2-nonenal (HNE), has been implicated specifically as a causative factor for the initiation and/or progression of various diseases. To date, however, little is known about the proteins and their modification sites susceptible to "carbonyl stress" by this RCS, especially in the liver. Using chemoprecipitation based on a solid-phase hydrazine chemistry coupled with LC-MS/MS bottom-up approach and database searching, we identified several protein-HNE adducts in isolated rat liver mitochondria upon HNE exposure. The identification of selected major protein targets, such as the ATP synthase β-subunit, was further confirmed by immunoblotting and a gel-based approach in combination with LC-MS/MS. A network was also created based on the identified protein targets, which showed that the main protein interactions were associated with cell death, tumor morphology and drug metabolism, implicating the toxic nature of HNE in the liver mitoproteome. The functional consequence of carbonylation was illustrated by its detrimental impact on the activity of ATP synthase, a representative major mitochondrial protein target for HNE modifications.
Insights
Reactive carbonyl species like 4-hydroxy-2-nonenal (HNE) cause protein carbonylation. This study identifies HNE-modified proteins in rat liver mitochondria, revealing impacts on ATP synthase and cell death pathways.
Area of Science:
- Biochemistry
- Molecular Biology
- Toxicology
Background:
- Protein carbonylation is linked to various diseases.
- Reactive carbonyl species (RCS), such as 4-hydroxy-2-nonenal (HNE), are implicated in disease progression.
- Limited knowledge exists on proteins and modification sites susceptible to HNE-induced carbonyl stress, particularly in the liver.
Purpose of the Study:
- To identify specific protein targets of HNE modification in rat liver mitochondria.
- To investigate the functional consequences of HNE-induced protein carbonylation.
- To explore the biological networks associated with HNE-modified proteins in the liver mitoproteome.
Main Methods:
- Chemoprecipitation using solid-phase hydrazine chemistry.
- Liquid chromatography-tandem mass spectrometry (LC-MS/MS) for protein identification.
- Immunoblotting and gel-based approaches for validation.
- Bioinformatic network analysis of identified protein targets.
Main Results:
- Several protein-HNE adducts were identified in isolated rat liver mitochondria following HNE exposure.
- Major targets, including the ATP synthase β-subunit, were confirmed.
- Network analysis revealed associations with cell death, tumor morphology, and drug metabolism.
- HNE modification detrimentally impacted ATP synthase activity.
Conclusions:
- HNE-induced protein carbonylation affects key mitochondrial proteins in the liver.
- These modifications have functional consequences, impacting cellular processes like energy production.
- HNE plays a toxic role in the liver mitoproteome, contributing to pathophysiological conditions.
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