Acylation of Lens Proteins

晶状体蛋白的酰化

• 批准号: 9765327
• 负责人: Ram H Nagaraj
• 金额: $ 39.33万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9765327
• 关键词: Acetyl Coenzyme A; Acetylation; Acyl Coenzyme A; Acylation; Adopted; Advanced Glycosylation End Products; Age; Aging; Amino Acids; Biochemical; Biochemical Reaction; Blindness; Cataract; Cells; Chemicals; Crystallins; Cysteine; Data; Exhibits; Human; In Vitro; Lead; Lipids; Lysine; Malonyl Coenzyme A; Measures; Mediating; Modification; Molecular Chaperones; Mutation; N acylation; Oxides; Pathway interactions; Pattern; Post-Translational Protein Processing; Presbyopia; Proteins; Reaction; Sirtuins; Site; Solubility; Structure; Testing; Tissues; Transferase; Translating; Visual impairment; acyl group; age effect; age related; aged; alpha-Crystallins; amino group; base; case-based; chemical reaction; experimental study; glycation; improved; lens; lens transparency; novel therapeutics; prevent; propionyl-coenzyme A; protein crosslink; protein structure function; resilience; succinyl-coenzyme A; therapy development; water solubility

N-acylation of cytosolic proteins is a common occurrence in tissues, and it regulates protein structure and function. The major N-acylation types in cells are acetylation, propionylation, succinylation and malonylation, which are initiated by the enzymatic or non-enzymatic transfer of acetyl CoA, propionyl CoA, succinyl CoA and malonyl CoA to lysine residues, respectively. Lens proteins are long lived with little or no turnover, and thus, post-translational modifications (PTMs) accumulate with age. The lack of enzymatically active proteins, particularly in the core of the lens, prohibits the reversal of PTMs. Thus, if deacylases are weak in the lens, acylation is mostly a one-way reaction that leads to permanent modifications of lens proteins, which appears to be the case (based on our preliminary data). The effects of acylation on lens proteins remain largely unknown. As shown in our previous study, the acetylation makes α-crystallin a better chaperone. In addition, our preliminary data show that lens proteins are also propionylated, succinylated and malonylated and that acetylation and succinylation increase the thermal stability and solubility of the proteins. Thus, we hypothesize that acylation of lens proteins is a beneficial PTM that helps maintain the chaperone activity of α- crystallin as well as the stability and solubility of lens proteins during aging. We will test this hypothesis in three aims. In Aim 1, we will determine the major acylation sites in human lens α-crystallin by mass spectrometric analyses and determine the effect of age on these modifications. In Aim 2, we will determine effects of acylation on the structure and function of α-crystallin. In Aim 3, we will evaluate the effects of acylation on the stability and solubility of lens proteins and determine whether acylation can be used to improve the compromised resilience of aged human lenses.

胞质蛋白的N-酰化是组织中常见的，它调节蛋白质结构和 功能。细胞中主要的N-酰化类型是乙酰化，丙酰化，琥珀酰化和误导化， 由乙酰基CoA，丙二酮COA，琥珀酰COA的酶促转移或非酶转移引发的 分别为赖氨酸残留物。晶状体蛋白长期存在，几乎没有营业额，并且 因此，翻译后修饰（PTM）随着年龄的增长而积累。缺乏酶活性蛋白质， 特别是在镜头的核心中，禁止PTM的逆转。如果透镜中的脱酰酶很弱， 酰化主要是一种单向反应，导致晶状体蛋白的永久修饰，它出现 是这种情况（基于我们的初步数据）。酰化对晶状体蛋白的影响很大程度 未知。如我们先前的研究所示，乙酰化使α-晶状体蛋白成为更好的伴侣。此外， 我们的初步数据表明，晶状体蛋白也是丙酰化，琥珀酰化和误导的，并且 乙酰化和琥珀酰化增加了蛋白质的热稳定性和溶解度。那，我们 假设晶状体蛋白的酰化是一种有益的PTM，有助于维持α-的伴侣活性 衰老过程中晶状体蛋白的稳定性和溶解度。我们将在 三个目标。在AIM 1中，我们将通过质量确定人透镜α-晶状体中的主要酰基化位点 光谱分析并确定年龄对这些修饰的影响。在AIM 2中，我们将确定 酰化对α-晶蛋白结构和功能的影响。在AIM 3中，我们将评估 对晶状体蛋白的稳定性和溶解度的酰化，并确定是否可以使用酰基化 提高年迈的人类镜片的弹性。