Nuclear Magnetic Resonance Studies of Proteins

蛋白质的核磁共振研究

• 批准号: 6989407
• 负责人: ARTHUR G PALMER
• 金额: $ 30.5万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-08-01 至 2009-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6989407
• 关键词: Archaea; Escherichia coli; adenosine diphosphate; adenosine triphosphate; bacterial proteins; biophysics; chemical kinetics; enzyme activity; enzyme substrate complex; intermolecular interaction; membrane transport proteins; molecular dynamics; molecular site; nuclear magnetic resonance spectroscopy; nucleic acids; protein folding; protein structure function; ribonuclease H; site directed mutagenesis; species difference; structural biology; temperature; thermostability; transcription factor

DESCRIPTION (provided by applicant): Comprehensive experimental information on the essential contributions of intramolecular dynamics to biological functions of proteins is critical for biophysical theories of equilibrium properties, such as heat capacity and thermal stability; for mechanistic interpretations of kinetic processes, such as enzyme catalysis and ligand recognition; and for design of novel proteins and protein ligands, including pharmaceutical agents. This research project will use multidimensional NMR spectroscopy to address these fundamental issues. One long-term goal is to define the molecular determinants of stability and catalytic activity of the enzyme ribonuclease HI (RNase H) by comparing the structural, dynamical and enzymatic properties of homologous proteins derived from Escherichia coli and the extremely thermophilic bacterium Thermus thermophilus. The enzyme is distributed widely in prokaryotes and eukaryotes, and retroviral reverse transcriptase contains a C-terminal RNase H domain. Another long-term goal is to define the molecular determinants of ligand binding, including aspects of specificity, and allosterism, in DNA recognition by the yeast protein GCN4 and in nucleotide binding by the ATP-binding cassette (ABC) MJ1267 of the branched chain amino acid (LIV) transporter from Methanococcus jannaschii. GCN4 is the prototypical member of the bZip family of transcription activators. Motifs that recognize specific DNA sequences are ubiquitous components of proteins that regulate gene expression; consequently, explication of the molecular basis for recognition is critical for understanding normal biological function and pathology. GCN4 represents an example of induced fit molecular recognition through a disorder-order transition associated with DNA binding. The LIV transporter is a member of a diverse family of ABC transporters involved in numerous biological processes and diseases, including multidrug resistance and cystic fibrosis. ABC MJ1267 represents an example of selected-fit molecular recognition and allosteric transmission of conformational changes between remote sites in proteins.

描述（由申请人提供）：关于分子内动力学对蛋白质生物功能的重要贡献的综合实验信息对于平衡特性（例如热容量和热稳定性）的生物物理理论至关重要；用于动力学过程的机械解释，例如酶催化和配体识别；以及用于设计新型蛋白质和蛋白质配体，包括药剂。该研究项目将使用多维核磁共振波谱来解决这些基本问题。一个长期目标是通过比较源自大肠杆菌和极端嗜热细菌嗜热栖热菌的同源蛋白的结构、动力学和酶学特性，确定核糖核酸酶 HI (RNase H) 稳定性和催化活性的分子决定因素。该酶广泛分布于原核生物和真核生物中，逆转录病毒逆转录酶含有C端RNase H结构域。另一个长期目标是定义配体结合的分子决定因素，包括酵母蛋白 GCN4 的 DNA 识别和支链氨基的 ATP 结合盒 (ABC) MJ1267 的核苷酸结合的特异性和变构性方面。来自詹氏甲烷球菌的酸（LIV）转运蛋白。 GCN4 是转录激活因子 bZip 家族的典型成员。识别特定 DNA 序列的基序是调节基因表达的蛋白质中普遍存在的成分。因此，阐明识别的分子基础对于理解正常的生物功能和病理学至关重要。 GCN4 代表了通过与 DNA 结合相关的无序转换诱导拟合分子识别的示例。 LIV 转运蛋白是 ABC 转运蛋白不同家族的成员，参与多种生物过程和疾病，包括多药耐药性和囊性纤维化。 ABC MJ1267 代表了蛋白质中远程位点之间选择性配合分子识别和构象变化变构传递的一个例子。