RUI: Role and Formation of Tyrosine-Cysteine Protein Cofactors

RUI：酪氨酸-半胱氨酸蛋白辅因子的作用和形成

• 批准号: 1058391
• 负责人: David Benson
• 金额: $ 34.4万
• 依托单位: Calvin University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-01-15 至 2015-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1058391&HistoricalAwards=false
• 关键词: RUI; Role; Formation; Tyrosine; Cysteine

This award will investigate the ability of tyrosine and cysteine sidechains of proteins to crosslink through an arylalkylthioether bond (abbreviated as Tyr-Cys). Tyr-Cys crosslinks participate in oxidation catalysis and are formed by two proton-coupled oxidations. Metal-oxygen chemistry and the correct sidechain geometries are necessary for Tyr-Cys crosslink formation. Only four examples have been identified using X-ray crystallography that suggests limitations from the chemistry, sidechain geometries, or detection methods. A search of all protein structures in the protein databank showed many proteins with appropriate sidechain geometries for Tyr-Cys crosslink formation. Based on conditions for dityrosine formation, the chemistry for Tyr-Cys should be facile under biological conditions. Tyr-Cys identification by X-ray crystallography, however, requires careful analysis of high-resolution data. In order in better identify Tyr-Cys crosslinks a proteolysis and HPLC separation method will be employed using mass spectrometry and nuclear magnetic resonance detection. Cysteine dioxygnease contains a Tyr-Cys crosslink and will be examined for method validation. The three other proteins where Tyr-Cys have not been previously observed will be used to test if the combination of these detection methods and a previous protein databank search will expand the number of Tyr-Cys crosslinks simply examining static protein structure. This information, along with reexamining the function of Tyr-Cys in cystiene dioxygenase, will determine if the biological scope of Tyr-Cys containing proteins is significantly broader than currently thought.Broader impacts of this award come from the potential to expand the biological scope of the Tyr-Cys crosslink and parent crosslinked protein derived cofactors. Additionally, this award will use NSF-supported instrumentation and a newly renovated interdisciplinary laboratory. This work will prepare four undergraduate chemistry/biochemistry majors for graduate work in areas related to protein chemistry. This award will also provide indirect training for up to ten additional undergraduate researchers in a shared interdisciplinary research lab. Two high school students who are underrepresented minorities in the sciences will be mentored by the PI and co-PI in conjunction with ACS Project SEED.

该奖项将研究蛋白质的酪氨酸和半胱氨酸侧链通过芳基烷基thioether键交联的能力（缩写为Tyr-Cys）。 Tyr-Cys交叉链接参与氧化催化，并由两个质子耦合的氧化形成。对于Tyr-Cys的交联成是必需的，金属氧化学和正确的侧链几何形状是必需的。仅使用X射线晶体学鉴定了四个示例，该晶体晶体学表明了化学，侧链几何或检测方法的局限性。搜索蛋白质数据库中所有蛋白质结构的搜索显示许多蛋白质具有适当的Sidechain几何形状，用于Tyr-Cys的交联成。基于二乙胺形成的条件，在生物学条件下，Tyr-Cys的化学应容易。但是，通过X射线晶体学识别Tyr-Cys需要仔细分析高分辨率数据。为了更好地识别Tyr-Cys交联，将使用质谱和核磁共振检测来使用蛋白水解和HPLC分离方法。半胱氨酸二氧化酶包含一个Tyr-Cys的交叉链接，并将检查用于方法验证。先前未观察到的Tyr-Cys的其他三种蛋白质将用于测试这些检测方法的组合和先前的蛋白质数据库搜索是否会扩大简单检查静态蛋白质结构的Tyr-Cys交联的数量。该信息以及重新审查Tyr-Cys在氰基二氧酶中的功能，还将确定含有蛋白质的Tyr-Cys的生物学范围是否比当前想象的要广泛。此外，该奖项将使用NSF支持的仪器和新装修的跨学科实验室。这项工作将为与蛋白质化学有关的领域的研究生工作准备四个本科化学/生物化学专业。该奖项还将在共享的跨学科研究实验室中为多达十位本科研究人员提供间接培训。 PI和Co-Pi与ACS Project Seed一起指导了两名科学中代表性不足的高中生。