Regulation of hepatic lysine N-acetylation by cysteine proximity due to alcohol toxicity

酒精毒性导致的半胱氨酸接近对肝脏赖氨酸 N-乙酰化的调节

基本信息

批准号：
10752320
负责人：
Courtney McGinnis
金额：
$ 3.72万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-07-01 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10752320
关键词：
Acetaldehyde Acetates Acetylation Advanced Development Affect Age Alcohol consumption Alcoholic Liver Diseases Alcoholic liver damage Alcohols Amines Amino Acids Analytical Biochemistry Binding Biochemical Biochemistry Biological Assay Biological Markers Biophysics Carbon Cell Respiration Cessation of life Charge Chemistry Chronic Classification Clinical Consumption Cysteine Data Data Set Development Drug Metabolic Detoxication Early Diagnosis Enzymes Equilibrium Ethanol Metabolism Ethanol toxicity Evaluation Fellowship Goals Grant Hepatic Homeostasis Individual Iron Lead Liver Liver diseases Lysine Mediating Mentors Mentorship Metabolic Metals Mitochondria Modeling Modification Mus National Center for Advancing Translational Sciences National Institute on Alcohol Abuse and Alcoholism National Research Service Awards Nicotinamide adenine dinucleotide Outcome Oxidation-Reduction Pathogenicity Pathologic Pathology Pathway interactions Patients Post-Translational Protein Processing Preventive therapy Proteins Proteome Proteomics Reaction Recombinants Regulation Research Research Personnel Role Signal Transduction Sirtuins Site Stress Structure Sulfhydryl Compounds Supportive care Therapeutic Therapeutic Intervention Training United States National Institutes of Health Work Zinc alcohol consequences alcohol effect alcohol exposure bench to bedside chronic alcohol ingestion clinically relevant detection method enzyme activity feeding hepatoprotective improved in vitro activity in vivo insight interdisciplinary approach liver injury mouse model new therapeutic target novel oxidation prevent preventable death protein function protein structure proteomic signature regenerative therapy repaired response success therapeutic target

项目摘要

PROJECT SUMMARY The goal of this Ruth L. Kirschstein NRSA F31 fellowship is to develop the expertise of the applicant in pathogenic mechanisms of alcohol-induced post-translational modifications (PTMs). Alcohol-associated liver disease (ALD) is a leading cause of preventable mortality worldwide causing approximately 5.9% of deaths every year, yet lacks preventative or regenerative therapies. The liver is the primary site of alcohol detoxification generating products that negatively impact hepatic biochemistry. A major consequence of alcohol metabolism is the induction of lysine acetylation, a metabolically induced PTM. One poorly characterized factor is the disruption to the network of cysteine thiols involved in the cysteine proteome, which is crucial to enzyme activity, protein structure, and signaling. Models of early-stage ALD have been applied to proteomic studies to better understand the downstream effects of alcohol metabolism and elucidate perturbed pathways. Using a 6-week Lieber-DeCarli (LD) model, we quantified thiol redox changes in the hepatic proteome employing a novel click-chemistry-based nHPLC-MS/MS assay. With the same model, we examined the impact of chronic alcohol consumption on hepatic lysine N-acetylation through quantitative analysis via acetyl-IP followed by nHPLC-MS/MS. Briefly, these studies identified that alcohol feeding induced an overall reduced cysteine proteome and increased lysine acetylation. Recent studies have identified a biophysical spatial association between lysine and cysteine amino acids leading to direct interactions. Lysine acetylation is amplified when cysteine residues are within 15 Å, referred to as a cysteine-lysine pair (CysLys). Our work suggests the alcohol-induced increase in N-acetylation is partially due to the acetylation of the more reactive cysteine thiol followed by transfer to a nearby lysine, resulting in an SàN acetyl transfer reaction. The proposed training plan will investigate this relationship in a chronic LD model by integrating acetyl- and thiol redox OMIC analyses to evaluate pathologically relevant protein targets. Therefore, the central hypothesis of this F31 application is that protein CysLys pairs are a mechanism for increased lysine acetylation and cysteine redox sensitivity during alcohol toxicity and contributes to the pathology of ALD. Specific aim 1 will characterize hepatic proteomic signatures impacted by Lys acetylation and Cys redox changes due to chronic alcohol metabolism using a multi-layered integrated analysis. Specific aim 2 will define regulatory mechanisms of SàN acetyl transfer in specific proteins critical to the progression of ALD. These aims will be interrogated utilizing a multidisciplinary approach that will provide a thorough understanding of lysine acetylation and thiol redox signaling by regulating protein structure. Completion of these specific aims will support the applicant in becoming an independent researcher in the field of alcohol-induced metabolic dysregulation.

项目概要 Ruth L. Kirschstein NRSA F31 奖学金的目标是培养申请人在以下领域的专业知识：酒精诱导的翻译后修饰（PTM）的致病机制。疾病 (ALD) 是全球可预防死亡的主要原因，每年约造成 5.9% 的死亡年，但缺乏预防或再生疗法。肝脏是酒精解毒的主要场所。产生对肝脏生物化学产生负面影响的产物是酒精代谢的一个主要后果。赖氨酸乙酰化的诱导（一种代谢诱导的 PTM）的破坏是一个尚不清楚的因素。涉及半胱氨酸蛋白质组的半胱氨酸硫醇网络，这对酶活性、蛋白质至关重要早期 ALD 的结构和信号传导模型已应用于蛋白质组学研究，以更好地理解。使用为期 6 周的 Lieber-DeCarli 分析酒精代谢的下游影响并阐明受干扰的途径。 (LD) 模型，我们采用一种新颖的基于点击化学的方法量化了肝蛋白质组中硫醇氧化还原的变化使用相同的模型，我们研究了慢性饮酒对肝脏的影响。赖氨酸 N-乙酰化通过乙酰基-IP 进行定量分析，然后进行 nHPLC-MS/MS。发现酒精喂养会导致半胱氨酸蛋白质组总体减少和赖氨酸乙酰化增加。最近的研究发现赖氨酸和半胱氨酸氨基酸之间存在生物物理空间关联当半胱氨酸残基在 15 Å 以内时，赖氨酸乙酰化会被放大，称为 a 我们的工作表明酒精诱导的 N-乙酰化增加部分是由于。将更具反应性的半胱氨酸硫醇乙酰化，然后转移到附近的赖氨酸，从而产生 SàN 拟议的训练计划将通过慢性 LD 模型研究这种关系。整合乙酰基和硫醇氧化还原 OMIC 分析来评估病理相关的蛋白质靶点。该 F31 应用的中心假设是蛋白质 CysLys 对是赖氨酸增加的机制酒精中毒期间的乙酰化和半胱氨酸氧化还原敏感性，有助于 ALD 的病理学特异性。目标 1 将表征受 Lys 乙酰化和 Cys 氧化还原变化影响的肝脏蛋白质组特征使用多层综合分析的慢性酒精代谢将定义监管。特定蛋白质中 SàN 乙酰基转移的机制对 ALD 的进展至关重要。利用多学科方法进行询问，这将提供对赖氨酸乙酰化的透彻理解和硫醇氧化还原信号通过调节蛋白质结构来完成这些特定目标将支持申请人成为酒精引起的代谢失调领域的独立研究员。