STRUCTURAL REFINEMENT OF LEADZYME VIA DIPOLAR COUPLINGS

了解核酶的催化功能需要 关于分子动力学的信息。 铅依赖 核酶，或leadzyme，有几个特点，使其成为一个优秀的， 选择研究这个问题。 leadzyme的核心含有碱基 它们经历着不同时间的构象动力学 鳞片 使用13 C弛豫实验结合Lipari- 萨博形式主义，一个简单的模型，运动的基础内， 将开发leadzyme。 方法的最新发展 测量1H-13 C、1H-15 N和1H-1H偶极耦合将与 一种变体。 代替使用在磁场中排列的双胞 为了诱导前导酶的部分比对，丝状噬菌体Pf 1 将用于诱导leadzyme的部分排列。 PF 1已经 在帕迪实验室的初步工作中， 表示Pf 1易于比对寡核苷酸。 1H-13 C偶极 来自leadzyme核心的耦合将提供新的角度限制 将用于核心残基的结构改进。 的 - 用于芯内的基部的运动的模型的组合， 一个完善的核心模型将提供洞察的作用， leadzyme活性的构象动力学。 这也可能导致 为了更清楚地了解构象动力学在 核酶功能

Understanding the catalytic function of ribozymes will require information about the dynamics of the molecule. The lead-dependent ribozyme, or leadzyme, has several features that make it an excellent choice for examining this question. The leadzyme's core contains bases which are experiencing conformational dynamics of a variety of time- scales. Using 13C relaxation experiments in conjunction with Lipari- Szabo formalism, a simple model of the motions of bases within the leadzyme will be developed. The recent development of methods for measuring 1H-13C, 1H-15N, and 1H-1H dipolar couplings will be used with a variation. Instead of using bicelles that align in the magnetic field to induce partial alignment of the leadzyme, the filamentous phage Pf1 will be used to induce partial alignment of the leadzyme. Pf1 has been shown to align in a magnetic field and preliminary work in the Pardi lab indicates Pf1 readily aligns oligonucleotides. The 1H-13C dipolar couplings from the leadzyme's core will provide new angular restraints that will be used for structural refinement of the core residues. The combination of a model for the motions of the bases within the core and a refined model of the core will provide insight into the role of conformational dynamics in the leadzyme's activity. This may also lead to a clearer picture of the general role of conformational dynamics in ribozyme function.