Probing the Role of the N-Terminal Arm in EcoRI Endonuclease Structure and Function

德州农工大学的启动基金将支持Cynthia M. Dupureur教授的研究。本研究的主题是限制性内切酶的结构与功能的关系。研究的重点是EcoRI限制性内切酶。虽然酶的结构是已知的，但n端臂的位置和作用在晶体学上是不确定的。在本研究中，通过肽合成、光谱、金属离子结合实验和非共价重构实验，研究了所需的n端臂与酶的其余部分、金属离子和底物之间的关系。获得的信息有助于阐明这些反应组分之间的关系，以及澄清EcoRI活性的工作模型。这项研究将促进对限制性内切酶的基本理解，这对开发靶向核酸的化疗药物至关重要。

This starter grant award to Texas A&M will support the research of Professor Cynthia M. Dupureur. The theme of the research is the relationship of structure and function in restriction enzymes. Studies focus on EcoRI restriction endonuclease. While the structure of the enzyme is known, the position and thus role of the N-terminal arm is crystallographically undefined. In this research, the relationships among the required N-terminal arm and the rest of the enzyme, metal ions, and substrate are investigated using peptide synthesis, spectroscopy, metal ion binding assays, and non-covalent reconstitution experiments. Information obtained leads to elucidation of the relationships among these reaction components, as well as to clarification of working models of EcoRI activity. This research will advance the fundamental understanding of restriction enzymes essential for development of chemotherapeutics that target nucleic acids.