STRUCTURAL STUDIES OF PROTEIN RECOGNITION

蛋白质识别的结构研究

• 批准号: 3898357
• 负责人: CHRISTIN FREDERICK
• 金额: --
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3898357
• 关键词: DNA; DNA binding protein; Escherichia coli; X ray spectrometry; aminoacid tRNA ligase; antineoplastics; arsenic; bacterial proteins; bacterial virus; computer graphics /printing; conformation; crystallization; drug metabolism; enzyme structure; enzyme substrate complex; genetic manipulation; genetic operator element; histones; immunoglobulin structure; immunoglobulins; monoclonal antibody; nonhistone nucleoprotein; nucleic acid chemical synthesis; oligonucleotides; peptide chemical synthesis; protein engineering; protein structure function; synthetic peptide; transfer RNA; virus protein

The recognition and subsequent interactions between a protein and other molecules play essential roles in many fundamental biological processes. The overall objective of this project is to understand the molecular basis of how proteins recognize other molecules, including small molecule ligands as well as macromolecules such as nucleic acids and proteins, using single crystal x-ray diffraction studies. The rationale of the work here is that proteins may possess certain characteristic folding "motifs" for their substrate binding functions. Specifically, we propose to crystallize and determine the three dimensional structure of several proteins systems. These include 1) proteins that interact with DNA: E. coli Hu protein, Ike and fd gene 5 protein and E. coli Rep enzyme and their complexes with DNA oligonucleotides; 2) Aminoacyl tRNA synthetase fragments and their complexes with tRNA; 3) Fab fragments of antiarsonate immunoglobulins; 4) Synthetic peptides possessing DNA binding activity. We hope to crystallize these proteins with and without their respective substrates so that we can visualize the structure of the protein before and after complex formation. Detailed knowledge of the protein structure will allow us to assess the conformational changes in the protein molecule and to predict ways of altering existing proteins or designing new proteins of various functions. The proposed studies in this project involve collaboration with other principal investigators in the program project. By combining the knowledge of recombinant DNA methodology, monoclonal antibody technology, nucleotide and peptide chemistry and x-ray crystallography within the program project, we hope ultimately to achieve the long range goal of understanding the structure, interactions and function of proteins and of designing new proteins of improved or desired functions.

蛋白质之间的识别和随后的相互作用 和其他分子在许多基本的 生物过程。 该项目的总体目标是 了解蛋白质如何识别其他蛋白质的分子基础 分子，包括小分子配体以及 大分子如核酸和蛋白质，使用单个 晶体X射线衍射研究。 这里的工作原理 蛋白质可能具有某些特征性折叠 “基序”，用于其底物结合功能。 我们特别 建议结晶并确定三维 几种蛋白质系统的结构。 1）蛋白质 与DNA相互作用的基因：E。coli Hu蛋白、Ike和fd基因5 蛋白质和E. 大肠杆菌Rep酶及其与DNA的复合物 2）氨酰基tRNA合成酶片段和 抗胂酸Fab片段 免疫球蛋白; 4）具有DNA结合的合成肽 活动 我们希望能将这些蛋白质结晶， 因此我们可以看到 复合物形成前后的蛋白质。 详细 蛋白质结构的知识将使我们能够评估 蛋白质分子的构象变化，并预测 改变现有蛋白质或设计新蛋白质的方法 各种功能。 本项目中拟议的研究涉及与以下方面的合作： 项目的其他主要研究人员。 通过 结合重组DNA方法学的知识， 单克隆抗体技术、核苷酸和肽 化学和X射线晶体学在项目中， 我们希望最终能达到长远目标， 了解的结构，相互作用和功能 蛋白质和设计新的蛋白质的改进或所需的 功能协调发展的