QUANTITATION OF PROTEIN TURNOVER IN THE HUMAN LENS USING THE 14C BOMB-PULSE

使用 14C BOMB-脉冲对人晶状体中的蛋白质周转进行定量

• 批准号: 8362763
• 负责人: Bruce A Buchholz
• 金额: $ 25.75万
• 依托单位: UNIVERSITY OF CALIF-LAWRNC LVRMR NAT LAB
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-01 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8362763
• 关键词: Adult; Age; Attenuated; Carbon; Cataract; Cells; Crystalline Lens; Crystallins; Data; Diffusion; Funding; Grant; Homeostasis; Human; Individual; Label; Life; Maintenance; Measures; Molecular; National Center for Research Resources; Nature; Nuclear; Nuclear Proteins; Nuclear Weapon; Oceans; Pathology; Physiologic pulse; Principal Investigator; Production; Proteins; Radioactive; Research; Research Infrastructure; Resource Development; Resources; Sampling; Source; Testing; Tracer; Treaty; United States National Institutes of Health; accelerator mass spectrometry; age related; aged; cost; fiber cell; in vivo; lens; macromolecule; middle age; milligram; prevent; protein degradation; repaired; stable isotope; tool

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Studying synthesis and repair of macromolecules in healthy humans is difficult. Above ground testing of nuclear weapons from 1955-1963 produced a large "bomb-pulse" of 14CO2, which was quickly distributed around the globe and intrinsically labeled all exchangeable carbon in the biosphere. Since the Test Ban Treaty in 1963, the atmospheric 14C bomb-pulse has been decreasing exponentially with a mean life of ~ 16 years, not due to radioactive decay, but due to diffusion and equilibration with the oceans and biosphere. All living humans have been labeled, and the 14C concentration in all organic macromolecules can sensitively identify when they were synthesized. In reality, we are all subjects in a long-term quasi-stable isotope tracer study in which molecular synthesis can be dated through the use of accelerator mass spectrometry (AMS) to count individual 14C atoms in sub-milligram samples. We seek to utilize this effective molecular chronometer to establish, quantify, and identify the specific nature of protein turnover in healthy adult human eye lenses. Preliminary data from lenses age 60 and greater suggests that the crystallin proteins of aged nuclear fiber cells contain carbon significantly younger than the cells themselves. This serves as direct evidence of in vivo protein turnover in human nuclear fiber cells and verifies the controversial hypothesis that the human eye lens maintains homeostasis at its core (at least in part) by de novo protein production. We will measure carbon turnover in purified proteins from the nuclear core of lenses formed after the peak of the bomb-pulse plus some aged controls. Compared with the data gathered from older donors, dating protein incorporation in younger lenses will provide direct evidence if this rate is attenuated when we reach middle age. A better understanding of lens maintenance could provide a useful new tool to understand and ultimately prevent the most common form of lens pathology, age related nuclear (ARN) cataract.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 很难研究健康人类大分子的合成和修复。 从1955年至1963年开始对核武器进行地面测试产生了14CO2的大型“炸弹 - 脉冲”，该武器在全球范围内迅速分布，并在生物圈中固有地标记了所有可交换的碳。自1963年禁止测试禁令以来，大气14C炸弹脉冲的平均寿命约为16年，而不是由于放射性衰减，而是由于扩散和与海洋和生物圈的平衡所致。所有活人都被标记了，所有有机大分子中的14C浓度均可敏感地识别它们合成的何时。 实际上，我们都是长期准稳定的同位素示踪剂中的受试者，在该研究中，可以通过使用加速器质谱法（AMS）来计算亚毛皮样品中的单个14C原子，从而可以通过分子合成来记录分子合成。 我们寻求利用这种有效的分子计时仪来建立，量化和确定健康成人人眼镜蛋白更新的特定性质。来自60岁及60岁镜头的初步数据表明，老化的核纤维细胞的结晶蛋白含有比细胞本身明显年轻的碳。这是人体核纤维细胞中体内蛋白质更新的直接证据，并验证了人眼镜镜的稳态（至少部分）由从头蛋白产生的核心（至少部分）。我们将从炸弹脉冲峰和一些老年对照组后形成的透镜的核心核心中测量纯化蛋白质的碳更换。与从老年捐助者那里收集的数据相比，如果我们达到中年时，年轻镜头中的约会蛋白掺入将直接证据。更好地了解晶状体维护可以为理解并最终预防最常见的晶状体病理学，与年龄相关的核（ARN）白内障提供有用的新工具。