Evolution of the mechanism of peptidylglycine-alpha-amidating monooxygenase

肽基甘氨酸-α-酰胺化单加氧酶机制的演变

• 批准号: 1517522
• 负责人: L. Mario Amzel
• 金额: $ 102.87万
• 依托单位: Johns Hopkins University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-01 至 2019-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1517522&HistoricalAwards=false
• 关键词: Evolution; mechanism; peptidylglycine; alpha; amidating

Title: Evolution of the mechanism of peptidylglycine-alpha-amidating monooxygenase Peptides are very small proteins made up of short and precise sequences of amino acid building blocks. Many peptides serve as signaling agents such as hormones and neurotransmitters in complex multicellular organisms. After synthesis many of these signaling peptides require further modification for activation. In this project, the investigator will study the enzymes needed for a specific peptide modification from several distantly related organisms. The goal of the research is to identify different structural and chemical changes across the different organisms that resulted in the gain or the loss of different features of the enzyme. Determination of the mechanism of this important step in the modification of peptide signaling molecules and how the mechanism changes in different organisms will guide the development of new ways of producing synthetic modified peptides. The work will also lead to a better understanding of how different enzyme structures can give rise to improved efficiency or specificity for a chemical reaction. The laboratory of the PI will continue its efforts to bring the excitement of science to high school and undergraduate students with emphasis on including underrepresented minorities in the research programs and STEM education.Communication between different parts of an organism via secreted peptides is a widely distributed biological signaling mechanism. Many of these peptides require amidation at their carboxy terminus for full-activity. Such peptides are expressed as precursors that, after processing, contain a glycine residue as their last residue. Oxidative cleavage of the glycine N-C(alpha) bond produces a des-glycine amidated peptide and glyoxylate. Only one enzyme in the complete phylogenetic spectrum is known to catalyze this reaction: the bi-functional peptidylglycine-alpha-amidating monooxygenase. The reaction takes place in two stages, the hydroxylation of the glycine C(alpha) by peptidyl-glycine-alpha-hydroxylating monooxygenase, and cleavage of the N-C(alpha) bond by peptidylglycine-alpha-amidating lyase. This project will study the peptidylglycine-alpha-amidating monooxygenases from several distantly related organisms with the goal of determining when in evolution they appeared, while trying to identify the structural and chemical changes that resulted in the gain or the loss of different features of the enzyme not critical for the chemical reaction. Structural and biochemical information obtained by X-ray diffraction, small angle scattering and kinetic measurements of the proteins from evolutionarily distant organisms will be used in this work.

标题：肽基甘氨酸-α-酰胺化单加氧酶机制的演变 肽是非常小的蛋白质，由短而精确的氨基酸结构单元序列组成。 许多肽在复杂的多细胞生物体中充当信号剂，例如激素和神经递质。 合成后，许多这些信号肽需要进一步修饰才能激活。在这个项目中，研究人员将研究几种远缘相关生物体进行特定肽修饰所需的酶。 该研究的目标是确定不同生物体中不同的结构和化学变化，这些变化导致酶不同特征的获得或丧失。 确定肽信号分子修饰中这一重要步骤的机制以及该机制在不同生物体中如何变化将指导生产合成修饰肽的新方法的开发。 这项工作还将有助于更好地理解不同的酶结构如何提高化学反应的效率或特异性。 PI 实验室将继续努力为高中生和本科生带来科学的乐趣，重点是将代表性不足的少数群体纳入研究项目和 STEM 教育中。生物体不同部分之间通过分泌肽进行的交流是一种广泛分布的生物技术。信号机制。 许多这些肽需要在其羧基末端进行酰胺化才能发挥全部活性。 此类肽被表达为前体，在加工后，其最后的残基含有甘氨酸残基。甘氨酸 N-C(α) 键的氧化裂解产生脱甘氨酸酰胺化肽​​和乙醛酸盐。 在完整的系统发育谱中，只有一种酶已知可以催化该反应：双功能肽基甘氨酸-α-酰胺化单加氧酶。 该反应分两个阶段进行，肽基甘氨酸-α-羟基化单加氧酶对甘氨酸C(α)进行羟基化，以及肽基甘氨酸-α-酰胺化裂合酶对N-C(α)键进行裂解。该项目将研究来自几种亲缘关系较远的生物体的肽基甘氨酸-α-酰胺化单加氧酶，目的是确定它们在进化中何时出现，同时试图识别导致酶不同特征获得或丧失的结构和化学变化对于化学反应并不重要。这项工作将使用通过 X 射线衍射、小角散射和动力学测量获得的来自进化距离较远的生物体的蛋白质的结构和生化信息。