Acetylation-Mediated Post-Translational Modification of Pyruvate Dehydrogenase Plays a Critical Role in the Regulation of the Cellular Acetylome During Metabolic Stress
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Aishwarya Rajakumar1, Sarah Nguyen1, Nicole Ford1
1Division of Metabolic and Cardiovascular Sciences, Burnett School of Biomedical Sciences, College of Medicine, University of Central Florida, Orlando, FL 32827, USA.
Insights
Cardiac ischemia significantly reduces protein acetylation, impacting cellular metabolism and cytoskeleton organization. This study identifies key acetylation changes in proteins involved in energy production pathways during ischemic stress.
Area of Science:
- Biochemistry
- Cardiovascular Biology
- Cellular Stress Response
Background:
- Cardiac diseases are a leading global cause of death, often stemming from ischemic events that disrupt cellular homeostasis and metabolism.
- Ischemic cardiomyopathy poses a significant global health challenge, necessitating a deeper understanding of cellular responses to ischemia.
- Post-translational modifications, such as protein acetylation, play a crucial role in cellular function during ischemic conditions.
Purpose of the Study:
- To investigate the impact of ischemia on cellular acetylation levels in cardiomyocytes.
- To identify specific proteins and pathways affected by differential acetylation during ischemic stress.
- To elucidate the role of protein acetylation in cellular adaptation and dysfunction under ischemic conditions.
Main Methods:
- Establishment of an in vitro ischemia model using neonatal rat ventricular cardiomyocytes.
- Quantification of cellular acetylation levels via Western blotting and affinity chromatography.
- Identification and quantification of differentially acetylated proteins using liquid chromatography-mass spectrometry.
Main Results:
- Ischemic conditions led to a significant reduction in overall cellular acetylation compared to normoxic controls.
- Mass spectrometry identified 179 acetylation sites on proteins in ischemic cardiomyocytes, with 121 downregulated and 26 upregulated.
- Affected proteins are primarily involved in cellular metabolism, sarcomere structure, and motor activity, with notable changes in the pyruvate dehydrogenase complex.
- Ischemia impacted acetyl-CoA biosynthesis and the tricarboxylic acid cycle through deacetylation of associated proteins.
Conclusions:
- Differential protein acetylation during ischemia influences cellular metabolism and cytoskeleton organization.
- Understanding these acetylation changes offers potential therapeutic targets to mitigate ischemia's detrimental effects.
- This research provides insights into the dynamic regulation of protein function at specific sites during cellular stress.
Abstract:
Background: Cardiac diseases remain one of the leading causes of death globally, often linked to ischemic conditions that can affect cellular homeostasis and metabolism, which can lead to the development of cardiovascular dysfunction. Considering the effect of ischemic cardiomyopathy on the global population, it is vital to understand the impact of ischemia on cardiac cells and how ischemic conditions change different cellular functions through post-translational modification of cellular proteins. Methods: To understand the cellular function and fine-tuning during stress, we established an ischemia model using neonatal rat ventricular cardiomyocytes. Further, the level of cellular acetylation was determined by Western blotting and affinity chromatography coupled with liquid chromatography-mass spectroscopy. Results: Our study found that the level of cellular acetylation significantly reduced during ischemic conditions compared to normoxic conditions. Further, in mass spectroscopy data, 179 acetylation sites were identified in the proteins in ischemic cardiomyocytes. Among them, acetylation at 121 proteins was downregulated, and 26 proteins were upregulated compared to the control groups. Differentially, acetylated proteins are mainly involved in cellular metabolism, sarcomere structure, and motor activity. Additionally, a protein enrichment study identified that the ischemic condition impacted two major biological pathways: the acetyl-CoA biosynthesis process from pyruvate and the tricarboxylic acid cycle by deacetylation of the associated proteins. Moreover, most differential acetylation was found in the protein pyruvate dehydrogenase complex. Conclusions: Understanding the differential acetylation of cellular protein during ischemia may help to protect against the harmful effect of ischemia on cellular metabolism and cytoskeleton organization. Additionally, our study can help to understand the fine-tuning of proteins at different sites during ischemia.
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