STRUCTURAL REFINEMENT OF LEADZYME VIA DIPOLAR COUPLINGS

了解核酶的催化功能将需要 关于分子动力学的信息。铅依赖者 核酶，或铅酶，有几个特点，使其成为一种优秀的 考查这个问题的选择。铅酶的核心含有碱基 它们正经历着不同时间的构象动力学- 比例。将13C弛豫实验与Lipari- Szabo形式主义，一个简单的底座运动模型 将开发铅酶。近几年来国内外研究方法的进展 测量1H-13C、1H-15N和1H-1H偶极联轴器将用于 一种变种。而不是使用在磁场中对齐的双柱体 为了诱导丝状噬菌体PF1的部分序列比对 将被用来诱导铅酶的部分排列。PF1已经成为 显示了在磁场中排列，并在Pardi实验室进行了初步工作 表明PF1很容易与寡核苷酸对齐。1H-13C偶极 铅酶核心的联结将提供新的角度限制 这将用于核心残基的结构精炼。这个 核心内底座运动的模型和 核心的精炼模型将提供对以下角色的洞察 铅酶活性中的构象动力学。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.