RUI: Atranyl-Nucleosides as Ribozyme Probes

RUI：Atranyl 核苷作为核酶探针

• 批准号: 0135279
• 负责人: Edward Fenlon
• 金额: $ 9.6万
• 依托单位: Xavier University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-05-01 至 2003-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0135279&HistoricalAwards=false
• 关键词: RUI; Atranyl; Nucleosides; Ribozyme; Probes

Current models for the catalytic mechanisms of protein enzymes are far more advancedthan those for RNA enzymes (ribozymes). This project will contribute to understanding how thecatalytic mechanisms of ribozymes compare with those of protein enzymes. The specificobjectives of this project are to: (1) synthesize atranyl-nucleosides 1-3; (2) characterize anddetermine the aqueous stability of atranyl-nucleosides; (3) use atranyl-nucleosides as transitionstate analogues (TSAs) to study the hammerhead and lead-dependent (leadzyme) ribozymes viacollaborations; and (4) train undergraduate students in synthetic organic techniques inpreparation for graduate study and/or careers in science. TSAs are useful for enzyme studies;however, suitable TSAs for phosphoryl transfer reactions are lacking. Compounds 1-3 have anatrane moiety incorporated into a ribonucleoside. The atrane contains a trigonal bipyramidalsilicon, germanium, tin, or titanium atom; and thus, 1-3 mimic the oxyphosphorane transitionstate of the ribozyme-catalyzed reaction. Compound 2 is a protected phosphoramidite thatcontains uridine and deoxyadenosine connected through the atrane moiety. Several syntheses of2 starting from 5'-O-DMT-2'-amino-2'-deoxyuridine will be evaluated. Prof. William G. Scott ofUC-Santa Cruz will incorporate 2 into the hammerhead substrate by solid-phase synthesis toobtain a hammerhead ribozyme-TSA crystal structure. Prof. Arthur Pardi of the University ofColorado-Boulder will similarly determine the structure in solution using NMR spectroscopy.Analogous approaches will be taken for leadzyme structural studies following the synthesis ofphosphoramidite 3. These experiments will elucidate the magnitude of conformationalrearrangement required to reach the transition state in each of these small RNA catalysts. Thisresearch will also help identify the role of specific RNA functional groups and metal cations incatalysis. This knowledge is paramount for understanding the pre-biotic "RNA world" and howour current life forms evolved from it.

蛋白质酶催化机制的当前模型的速度要高得多，而RNA酶（核酶）的模型要高得多。该项目将有助于理解核酶的催化机制与蛋白质酶相比。该项目的特定对象是：（1）合成丁那酰核苷1-3； （2）表征并确定苯基核苷的水稳定性； （3）使用丙酰基核苷作为过渡州类似物（TSA）来研究锤头和铅依赖性（Leadzyme）核酶的核糖因杀伤性； （4）培训本科生的合成有机技术，用于研究生学习和/或科学职业。 TSA可用于酶研究；但是，缺乏适合磷酸化转移反应的TSA。化合物1-3的Anatrane部分掺入核糖核苷中。载体中包含三角形双锥子，锗，锡或钛原子。因此，1-3模拟核酶催化反应的羟磷酸烷转变状态。化合物2是一种受保护的磷酰胺，可连接通过胸膜部分连接的尿苷和脱氧腺苷。将评估从5'-O-DMT-2'-Amino-2'-脱氧尿苷的几个合成2的合成。 William G. Scott Ofuc-Santa Cruz教授将通过固相合成toObtain hammerhead核酶-TSA晶体结构将2种在锤头底物中。 Ofcolorado-Boulder的Arthur Pardi教授将类似地确定使用NMR光谱法进行溶液中的结构。在合成磷酸氧化岩的合成后，将采取销售方法进行铅酶结构研究。这些实验将阐明所需的构象良性果实型在每个小范围的过渡状态所需的型号的型号。本研究还将有助于确定特定的RNA官能团和金属阳离子的作用。这些知识对于理解益生元的“ RNA世界”和豪库当前的生命形式至关重要。