Probes for Studies of Nucleotide-Binding Proteins

用于研究核苷酸结合蛋白的探针

• 批准号: 6806316
• 负责人: MICHAEL GROZIAK
• 金额: $ 20.28万
• 依托单位: CALIFORNIA STATE UNIVERSITY HAYWARD
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-07-08 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6806316
• 关键词: X ray crystallography; analog; binding proteins; biomimetics; biotechnology; chemical bond; chemical structure; chemical synthesis; conformation; molecular probes; nuclear magnetic resonance spectroscopy; nucleotides; protein structure function; stereoisomer

DESCRIPTION (provided by applicant): This project focuses on the development of a novel class of transglycosidically tethered 5'-nucleotides by synthesis and physicochemical evaluation. A single general synthetic strategy for the tethering of any 5'-nucleotide of interest has already been developed. It was used successfully to prepare the first two tethered 5'-nucleotide targets. The specific nature of this new tether and its mode of attachment to a nucleotide framework allow an unprecedented degree of 5'-nucleotide biomimicry according to the conformational disposition of the protein-bound ligands in the Protein Data Bank. The tethered 5'-nucleotides are for all practical purposes chemically identical to their natural counterparts but are restricted to a limited set of spatial conformations. Further development will confirm their highly biomimetic features and demonstrate their utility to biomedical investigations. Synthesis of a variety of carefully selected targets, and extensive characterization by NMR spectroscopy and X-ray crystallography are the specific activities to be undertaken in this work. Conformational restriction can generate valuable probes of protein-ligand structure-function relationships. Just two bond rotations determine most of the molecular topography of 5'-nucleotides: the C4'-C5' bond and the glycosidic one (C1'-N1 for pyrimidines and C1'-N9 for purines), and these are restricted simultaneously by the new tether. The desirable features of a transglycosidic nucleotide-tethering motif include restriction of the glycosidic bond in an anti conformation, preservation of the biorecognition-critical hydrogen bonding sites of the heterocyclic aglycon, retention of the functionalizable carbohydrate hydroxyl groups, and a design that can be used on both the pyrimidine and the purine nucleoside frameworks. The tethering motif featured in this proposal meets all these criteria, and future use of the tethered targets could contribute to an understanding of conformation/binding/function relationships in many biochemically important 5'-nucleotide-binding proteins.

描述（由申请人提供）：该项目着重于通过合成和理化评估的新型跨糖化束缚5'-核苷酸的开发。已经开发了一种束缚任何5'-核苷酸的一般合成策略。它被成功地用于准备前两个束缚的5'-核苷酸靶标。根据蛋白质数据库中蛋白质结合的配体的构象处置，这种新系列的特定性质及其对核苷酸框架的附着模式允许5'-核苷酸的仿生型。束缚的5'-核苷酸在化学上与其自然对应物相同，但仅限于有限的空间构象。进一步的发展将确认其高度仿生的特征，并证明其对生物医学研究的效用。在这项工作中要进行的特定活性是多种精心选择的目标的合成，以及NMR光谱和X射线晶体学的广泛表征。 构象限制可以生成蛋白质配体结构函数关系的有价值的探针。只有两个键旋转决定了5'-核苷酸的大部分分子形态：C4'-C5'键和糖苷的键（嘧啶的C1'-N1，purimidines的C1'-N1，purines的C1'-N9），这些粘合剂是由新的Tether同时限制的。克糖苷核苷酸 - 螺旋序的理想特征包括在抗构象中限制糖苷键，保留生物识别 - 识别 - 识别氢键键合的位点，保留杂型敏捷的杂环敏捷位点，可以保留可官能化的核苷和设计的纯化核氢和设计，并在拟南芥中均可使用碳水化合物和设计。框架。该提案中符合所有这些标准的束缚主题符合所有这些标准，并且未来对束缚目标的使用可能有助于理解许多生物化学重要的5'-核苷酸结合蛋白中的构象/结合/功能关系。