NUCLEAR MAGNETIC RESONANCE--NEW METHODS AND MOLECULAR STRUCTURE DETERMINATION

核磁共振--分子结构测定的新方法

• 批准号: 6432089
• 负责人: Ad - Bax
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6432089
• 关键词: biomagnetism measurement; chemical binding; intermolecular interaction; magnetism; method development; nuclear magnetic resonance spectroscopy; protease inhibitor; protein structure; solvents; structural biology; water solution

We have continued our study of biological macromolecules dissolved in a very dilute, nematic liquid crystalline (LC) phase. Study of chemical shift aniostropy (CSA) of proton, carbon and nitrogen backbone atoms in the protein ubiquitin, dissolved in a LC medium, revealed distinct correlations between CSA and protein secondary structure. A method has been developed that quantitatively predicts alignment of macromolecules of known shape for the case where the LC particles are nearly neutral. This information confirmed the monomeric form of the anti-HIV protein cyanovirin-N in solution. Methods have been developed that permit measurement of dipolar couplings in slowly tumbling macromolecules. We have shown that in favorable cases it is possible to define the three-dimensional structure of a protein backbone solely on the basis of its dipolar couplings, provided that measurements can be performed in two different LC media. This development holds potential to significantly decrease the time required for protein structure determination. Dipolar couplings were used for determination of the structure of the RecA binding protein DinI, and for obtaining a high-resolution structure of the DNA dodecamer d(CGCGAATTCGCG)2.

我们继续研究溶解在非常稀释的列液晶（LC）相的生物大分子。 溶解在LC培养基中的蛋白质泛素中质子，碳和氮骨链原子的化学移位障碍物（CSA）的研究表明，CSA与蛋白质的二级结构之间存在明显的相关性。 已经开发了一种方法，该方法可以定量预测LC颗粒几乎中性的情况下已知形状的大分子的比对。该信息证实了溶液中抗HIV蛋白Cyanovirin-N的单体形式。 已经开发了允许在缓慢翻滚的大分子中测量偶极耦合的方法。 我们已经表明，在有利的情况下，只要可以在两种不同的LC介质中进行测量，就可以仅根据其偶极耦合来定义蛋白质主链的三维结构。这种发展具有显着减少蛋白质结构确定所需的时间的潜力。 偶极耦合用于确定RECA结合蛋白DINI的结构，并获得DNA DodeCamer D（CGCGAATTCGCG）的高分辨率结构2。