Molecular mechanisms of action of macrolide antibiotics

大环内酯类抗生素的分子作用机制

• 批准号: 8640960
• 负责人: ALEXANDER S MANKIN
• 金额: $ 30.31万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-04-01 至 2017-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8640960
• 关键词: Address; Affect; Anti-Bacterial Agents; Antibiotics; Bacterial Infections; Binding; Binding Sites; Biochemical; Bypass; Cells; Development; Genomics; Gram-Negative Bacteria; In Vitro; Ligand Binding; Macrolide Antibiotics; Macrolide-resistance; Macrolides; Medical; Modeling; Molecular; Molecular Mechanisms of Action; N-terminal; Nature; Peptides; Pharmaceutical Preparations; Physiological; Property; Protein Biosynthesis; Protein Synthesis Inhibitors; Proteins; Proteomics; Research; Resistance; Ribosomes; Site; Staging; Structure; System; Techniques; Translations; Variant; cell growth; drug development; evidence base; genome-wide; in vivo; innovation; novel; novel strategies; polypeptide; public health relevance

DESCRIPTION (provided by applicant): Macrolide antibiotics inhibit cell growth by interfering with protein synthesis. These drugs bind in the nascent peptide exit tunnel and, according to the widely accepted view, inhibit synthesis of all cellular proteins at the early rounds of translation In contrast to this conventional model of macrolide action, our preliminary studies showed that treatment of Gram-positive and Gram-negative bacteria with macrolides allows for continued translation of a defined subset of proteins. Further, the ability of the protein to evade inhibitio is determined by its N-terminal sequence which can bypass the antibiotic in the exit tunnel without displacing the drug from its binding site. After the initial bypass, translation of some proteins cn continue until their completion, whereas synthesis of some polypeptides can be arrested at later stages. Both of these effects depend on the structure of the antibiotic. In spite of the functional and medical significance of these phenomena, the molecular mechanisms underlying the ability of the protein to evade inhibition and the requirements for the drug-induced translation arrest are unknown and will be addressed in this project. Whole-cell proteomics will be used to comprehensively characterize proteins whose translation continues in the presence of the antibiotic. The highly-innovative technique of ribosome profiling will provide genome-wide information of the sites of drug-dependent 'late' translation arrest. The whole cell-studies will b followed by biochemical characterization of molecular mechanisms of bypass and arrest carried out in a cell-free translation system. Finally, the correlation between the structure of the antibiotic bound in the ribosomal exit tunnel and the spectrum of proteins synthesized in antibiotic-treated cells will be analyzed and physiological consequences of the variation in the composition of the resistome will be examined. The anticipated findings should significantly expand the understanding of the general mode of action of clinically-important macrolide antibacterials and open new venues for development of protein synthesis inhibitors with superior antibiotic properties.

描述（由申请人提供）：大环内酯类抗生素通过干扰蛋白质合成来抑制细胞的生长。这些药物在新生肽出口隧道中结合，根据被广泛接受的观点，在翻译早期抑制所有细胞蛋白的合成与这种传统的大月croadire作用模型相反，我们的初步研究表明，将革兰氏阳性和革兰氏阳性细菌与多果仁蛋白的蛋白质的蛋白质相比。此外，蛋白质逃避抑制剂的能力取决于其N末端序列，该序列可以绕过出口隧道中的抗生素而不会从其结合位点移位。在初始旁路之后，某些蛋白质CN的翻译一直持续到完成为止，而某些多肽的合成可以在以后的阶段停止。这两种作用都取决于抗生素的结构。尽管有功能 和这些现象的医学意义，即蛋白质逃避抑制作用和对药物诱导的翻译停滞能力的基础的分子机制尚不清楚，并且将在该项目中解决。全细胞蛋白质组学将用于全面表征其在抗生素存在下继续进行的蛋白质。核糖体分析的高度创新技术将提供全基因组依赖性“晚期”翻译停滞所在的信息。整个细胞研究将B随后进行生化表征，对在无细胞翻译系统中进行的旁路和停滞的分子机制进行了生化表征。最后，将分析核糖体出口隧道中结合的抗生素结构与抗生素处理细胞中合成的蛋白质光谱之间的相关性，并将检查抗抗性组组成中变异的生理后果。预期的发现应显着扩展对临床至关重要的大花环抗菌物的一般作用模式的理解，并开放具有优质抗生素特性的蛋白质合成抑制剂的开放新场所。