NUCLEAR MAGNETIC RESONANCE--NEW METHODS AND MOLECULAR STRUCTURE DETERMINATION

我们继续我们的研究，生物大分子溶解在一个非常稀，液晶（LC）相。对原有的基于磷脂的培养基进行了各种改进，将原来的5.5-7.5 pH范围扩大到2-11，温度范围从原来的30- 40 C增加到20- 50 C，这使得该技术适用于大多数感兴趣的系统。我们已经表明，在LC相偶极耦合的测量提供了非常精确的结构信息，这使得表征的相对，振动平均有效的核间距离和角度。我们已经表明，偶极耦合的测量可以提供一种快速的手段，用于评估普遍的解决方案的结构和结构模型之间的协议，也开发了方法，用于提高从NMR数据计算的结构的准确性。这项工作的大部分集中在一个小的模型蛋白，泛素，这已经很好地表征了晶体学和以前的NMR研究。该方法也正在各种其他系统上进行测试，包括DNA十二聚体，人类T细胞受体的V-α结构域和DinI，一种参与关闭SOS反应的蛋白质。- 核磁共振，蛋白质结构，液晶，胶束，双胞，磁取向

We have continued our study of biological macromolecules dissolved in a very dilute, nematic liquid crystalline (LC)phase. Various improvements to the original phospho lipid based medium have been made which extend the original 5.5-7.5 pH range to 2-11, and the temperature range which originally was 30-40Chas been increased to 20-50C, which makes the technology applicable to the majority of systems of interest. We have shown that measurement of dipolar couplings in the LC phase provides extremely precise structural information, which allows characterization of the relative, libration-averaged effective internuclear distances and angles. We have shown that measurement of dipolar couplings can provide a rapid means for evaluating the agreement between the prevalent solution structure and a structural model, and have also developed methods for improving the accuracy of structures calculated from NMR data. Most of this work has focused on a small model protein, ubiquitin, which has been well characterized by crystallographic and previous NMR studies. The methodology is also being tested on a variety of other systems, including a DNA dodecamer, the V-alpha domain of the human T-cell receptor, and DinI, a protein involved in shutting down the SOS response. - NMR, protein structure, liquid crystal, micelle, bicelle, magnetic alignment