X-ray Structural Studies of Proteins

蛋白质的 X 射线结构研究

• 批准号: 6572271
• 负责人: BRIAN W MATTHEWS
• 金额: $ 25.42万
• 依托单位: UNIVERSITY OF OREGON
• 依托单位国家: 美国
• 财政年份: 1979
• 资助国家: 美国
• 起止时间: 1979-01-01 至 2007-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6572271
• 关键词: DNA; DNA repair; DNA replication; Escherichia coli; RNA binding protein; X ray; X ray crystallography; adenosinetriphosphatase; aminopeptidase; beta galactosidase; binding proteins; bioimaging /biomedical imaging; crystallization; enzyme mechanism; enzyme structure; exonuclease; gene expression; genetic recombination; intermolecular interaction; neurogenesis; protein structure function; protein transport; site directed mutagenesis; structural biology

DESCRIPTION (provided by applicant): A major objective of this application is to obtain a better understanding of the interactions between nucleic acids and proteins and of related biological activities including the regulation of gene expression and the mechanisms of DNA replication, transcription and recombination. Toward this end crystallographic, biochemical and mutagenic studies will be pursued of a number of DNA- and RNA-interacting proteins including the exonuclease from phage lambda, exonuclease I from E. coli, the biotin repressor from E. coli, RNA splicing proteins from chloroplasts, proteins involved in asymmetric cell division in Drosophila and RNA-interacting DEAD box proteins. Studies will also be continued of E. coli beta-galactosidase, E. coil F1 ATPase and E. coli methionine aminopeptidase. We also hope to use the beta-galactosidase system to better understand the changes in protein structures that occur on flash-freezing and, at the same time, to develop systematic methods to avoid crystal cracking and other types of damage that often frustrate the use of this technique. The way in which proteins interact with DNA is fundamental to the correct regulation and growth of every living cell. Many human diseases have their origins in a breakdown in these basic control processes. Studies of the sort described here provide fundamental insights into the way in which proteins recognize their target sites on DNA and exert their control functions. There is every reason to expect that understandings derived from such systems in simple organisms will be relevant to human health. Knowledge of the three-dimensional structure of E. coli beta-galactosidase is expected to lead to improvements in a number of the assays that are based on the unique properties of the enzyme.

描述（由申请人提供）：本申请的主要目的是更好地理解核酸和蛋白质之间的相互作用以及相关的生物活性，包括基因表达的调节以及DNA复制、转录和重组的机制。 为此，将对许多 DNA 和 RNA 相互作用蛋白进行晶体学、生化和诱变研究，包括来自 lambda 噬菌体的核酸外切酶、来自大肠杆菌的核酸外切酶 I、来自大肠杆菌的生物素阻遏物、来自叶绿体的 RNA 剪接蛋白、参与果蝇不对称细胞分裂的蛋白质和 RNA 相互作用的 DEAD box 蛋白质。还将继续对大肠杆菌 β-半乳糖苷酶、大肠杆菌 F1 ATP 酶和大肠杆菌甲硫氨酸氨基肽酶进行研究。 我们还希望利用β-半乳糖苷酶系统更好地了解速冻时蛋白质结构的变化，同时开发系统方法以避免晶体破裂和其他类型的损坏，这些损坏通常会阻碍使用这项技术。 蛋白质与 DNA 相互作用的方式对于每个活细胞的正确调节和生长至关重要。许多人类疾病的根源在于这些基本控制过程的崩溃。本文描述的此类研究为蛋白质识别 DNA 上的靶位点并发挥其控制功能的方式提供了基本见解。我们有充分的理由相信，从简单生物体中的此类系统中获得的理解将与人类健康相关。了解大肠杆菌 β-半乳糖苷酶的三维结构预计将改进许多基于该酶独特性质的测定方法。