Crystallographic Studies on Ribosomal Particles

核糖体颗粒的晶体学研究

基本信息

批准号：
8135863
负责人：
ADA E YONATH
金额：
$ 16.2万
依托单位：
WEIZMANN INSTITUTE OF SCIENCE
依托单位国家：
美国
项目类别：
财政年份：
1985
资助国家：
美国
起止时间：
1985-08-01 至 2011-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8135863
关键词：
Amino Acids Antibiotics Archaea Base Pairing Binding Biochemical Biochemical Genetics Budgets Categories Cells Clinical effectiveness Complex Confusion Crystallography Data Drug Design Drug resistance pathway Effectiveness Elements Engineering Eubacterium Eukaryota Evolution Family Freezing Frequencies Genetic Code Genetic Models Goals Knowledge Length Life Ligands Light Maps Methods Modeling Molecular Molecular Chaperones Mutagenesis Mutate Mutation Mycobacterium smegmatis Nucleoproteins Nucleotides Organelles Oxazolidinones Peptides Peptidyltransferase Pharmaceutical Preparations Phase Positioning Attribute Predisposition Process Property Protein Biosynthesis Proteins RNA Research Personnel Resistance Resolution Ribosomes Site Species Specificity Structure System Temperature Therapeutic Transfer RNA Translations X ray diffraction analysis X-Ray Diffraction analog base cell growth regulation clinically relevant cryogenics design electron density improved inhibitor/antagonist insight mutant novel particle pathogen pleuromutilin polymerization programs protein folding rRNA Operon resistance mechanism stem synchrotron radiation telithromycin three dimensional structure tool

项目摘要

DESCRIPTION (provided by applicant): The long-term goal of our project has been and is shedding light on the molecular mechanisms involved in protein biosynthesis. Ribosomes, the universal cell organelles facilitating the translation of the genetic code into proteins, are nucleoprotein assemblies (2.3 mDa approximately 4500 RNA nucleotides and approximately 55 proteins) built of two subunits of unequal size, which associate upon the initiation of protein biosynthesis. The immediate objectives of this proposal are: (a) elucidating the detailed mechanisms involved in peptide bond formation, amino acid polymerization, peptide bond formation, translocation, tRNA release, nascent protein progression, elongation arrest and initial steps towards folding. For these studies carefully designed complexes capturing ribosomal particles at defined functional states, are being prepared, based on the high- resolution structures of the two eubacterial ribosomal subunits determined by us. Examples are complexes of substrate analogs, functional ligands, inhibitors, and non-ribosomal compounds relevant to protein biosynthesis, such as native and mutated trigger factor; (b) Determining the parameters acquiring the effectiveness of ribosomal antibiotics, by careful analysis of the distinction between mere binding and inhibitory action, based on antibiotics binding modes to ribosomes of authentic pathogens, of eubacteria serving as pathogen models, and of archaea resembling eukaryotes; (c) Further advance towards understanding mechanisms attaining resistance to ribosomal antibiotics, by elucidating the structural bases of resistance mechanisms developed against traditional as well as advanced ribosomal antibiotics. Methods: High resolution X-ray diffraction data are being collected at cryogenic temperatures from flash- frozen crystals of complexes of functionally active ribosomes, using high brilliance synchrotron radiation. Phases are being determined by a MIRAS, molecular replacement and crystal averaging. The resulting electron density maps are being interpreted interactively and the positions and orientations of the ligands and/or antibiotics are being compared to available biochemical and mutagenesis data. The significance of ribosomal crystallography stems from its potential to illuminate the mechanism of a fundamental life process, protein biosynthesis, as well as to reveal modes of action of antibiotics targeting ribosomes. These studies have already illuminated principles of drug selectivity, and provided significant basic knowledge of possible pathways of drug resistance, hence paving the way to improve therapeutic properties.

描述（由申请人提供）：我们项目的长期目标已经并且正在阐明蛋白质生物合成所涉及的分子机制。核糖体是促进遗传密码转化为蛋白质的通用细胞细胞器，是核蛋白组件（2.3 MDA约4500个RNA核苷酸和大约55个蛋白质），由两个不等大小的亚基建立，它们与蛋白质生物合成的起始相关。该提案的直接目标是：（a）阐明参与肽键形成，氨基酸聚合，肽键形成，易位，tRNA释放，新生蛋白进展，伸长延长和折叠的初步步骤的详细机制。对于这些研究，根据我们确定的两个长杆菌核糖体亚基的高分辨率结构，正在制备仔细设计的复合物，从而制备了在定义的功能状态下捕获核糖体颗粒。例子是与蛋白质生物合成相关的底物类似物，功能性配体，抑制剂和非核糖体化合物的复合物，例如天然和突变的触发因素；（b）通过仔细分析基于与真实病原体的核糖体的抗生素结合模式，将核糖体抗生素的有效性仔细分析，从而获得核糖体抗生素的有效性，从而获得抗生素的结合模式，这些模式可作为病原体模型和古细菌的厄生菌模型以及类似于厄尔纳西亚蛋白的核心；（c）通过阐明针对传统和晚期核糖体抗生素开发的耐药机制的结构碱基，进一步迈向理解对核糖体抗生素抗性的机制。方法：使用高光辉的同步辐射，正在从功能活性核糖体的复合物中收集高分辨率X射线衍射数据。相位由MIRAS，分子替代和晶体平均确定。所得的电子密度图正在交互作用，并将配体和/或抗生素的位置和方向与可用的生化和诱变数据进行比较。核糖体晶体学的重要性源于其阐明基本生命过程，蛋白质生物合成的机制，以及揭示靶向核糖体抗生素的作用模式。这些研究已经阐明了药物选择性的原则，并为耐药性的可能途径提供了重要的基本知识，从而为改善治疗特性铺平了道路。

项目成果

期刊论文数量（85）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Approaching the molecular structure of ribosomes.

接近核糖体的分子结构。

DOI：
10.1016/0301-4622(88)87021-2
发表时间：
1988
期刊：
Biophysical chemistry
影响因子：
3.8
作者：
Yonath,A;Wittmann,HG
通讯作者：
Wittmann,HG

Crystallographic studies on the ribosome, a large macromolecular assembly exhibiting severe nonisomorphism, extreme beam sensitivity and no internal symmetry.

对核糖体的晶体学研究，核糖体是一种大分子组装体，表现出严重的非同构性、极高的光束敏感性和无内部对称性。

DOI：
10.1107/s010876739800991x
发表时间：
1998
期刊：
Acta crystallographica. Section A, Foundations of crystallography
影响因子：
0
作者：
Yonath,A;Harms,J;Hansen,HA;Bashan,A;Schlünzen,F;Levin,I;Koelln,I;Tocilj,A;Agmon,I;Peretz,M;Bartels,H;Bennett,WS;Krumbholz,S;Janell,D;Weinstein,S;Auerbach,T;Avila,H;Piolleti,M;Morlang,S;Franceschi,F
通讯作者：
Franceschi,F

High-resolution structures of large ribosomal subunits from mesophilic eubacteria and halophilic archaea at various functional States.

不同功能状态下嗜温真细菌和嗜盐古菌的大核糖体亚基的高分辨率结构。