Synthesis and Use of Chiral Beta-Deuterated Amino Acids for NMR Studies of Protein Structure

手性 β-氘代氨基酸的合成和用于蛋白质结构 NMR 研究的用途

• 批准号: 9723642
• 负责人: Robert Curley
• 金额: $ 12万
• 依托单位: Ohio State University Research Foundation -DO NOT USE
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-09-01 至 2000-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9723642&HistoricalAwards=false
• 关键词: Synthesis; Use; Chiral; Beta; Deuterated

9723642 Curley The structure assignment of proteins as determined by modern NMR methods is greatly facilitated by access to appropriate regio- and stereoselectively labelled amino acids which can be incorporated into the proteins. Use of these labelled amino acids greatly increases experimental sensitivity and permits the use of isotope-edited NMR experiments which can provide specific structural information otherwise difficult to obtain. Of particular importance to assigning an accurate solution structure of a protein by NMR is stereospecific assignment of the amino acid side chain resonances. Useful for these assignments is access to amino acids with stereoselective substitution of deuteron for proton on the (-methylene groups of these amino acids with prochiral (-methylene protons. In addition, the effect of (-deuteration of the 13C relaxation rate will also facilitate C(carbon assignments, especially in large systems. A generalized, versatile, inexpensive method will be developed for the synthesis of virtually all the amino acids with prochiral (-methylene protons in a manner which will stereoselectively replace one of the prochiral protons with deuteron. These methods will also permit double labelling of the amino acid with 15N or 13C in a variety of patterns and can also be readily adapted to provide the natural amino acid L-isomer stereospecifically. Once obtained, these amino acids will be used to study biosynthesis and protein structure by NMR The function of proteins is dependent on their structure. Nuclear magnetic resonance (NMR) spectroscopy has become an important tool for determining the structure of proteins in solution. Proteins are composed of different amino acid building blocks which are bound together to make up the entire protein. The NMR study of proteins is made easier if the amino acids are labelled with nonradioactive, NMR-detectable isotopes. This project will develop general methods for the synthesis of these specially labelled amino acids. ***

9723642 Curley通过现代NMR方法确定的蛋白质的结构归属通过获得适当的区域和立体选择性标记的氨基酸而大大促进，所述氨基酸可以掺入蛋白质中。 使用这些标记的氨基酸大大增加了实验灵敏度，并允许使用同位素编辑的NMR实验，可以提供特定的结构信息，否则难以获得。 氨基酸侧链共振的立体特异性分配对于通过NMR分配蛋白质的准确溶液结构特别重要。 对于这些分配有用的是获得具有立体选择性的氘取代这些氨基酸的（-亚甲基上的质子的氨基酸，这些氨基酸具有前手性（-亚甲基质子。 此外，13 C弛豫速率的（-氘代的影响也将促进C（碳分配，特别是在大系统中。 一种通用的，通用的，廉价的方法将被开发用于合成几乎所有的氨基酸与前手性的α-亚甲基质子的方式，将立体选择性地取代一个前手性质子与氘核。 这些方法还允许用15 N或13 C以多种模式对氨基酸进行双标记，并且还可以容易地进行调整以立体特异性地提供天然氨基酸L-异构体。 一旦获得，这些氨基酸将被用于通过核磁共振研究生物合成和蛋白质结构 蛋白质的功能取决于其结构。 核磁共振（NMR）光谱已成为确定溶液中蛋白质结构的重要工具。 蛋白质由不同的氨基酸组成，这些氨基酸结合在一起构成整个蛋白质。 如果用非放射性的核磁共振可检测的同位素标记氨基酸，则蛋白质的核磁共振研究变得更容易。 该项目将开发合成这些特殊标记的氨基酸的一般方法。 ***