Hansen M R, Hanson P, Pardi A
a Department of Chemistry and Biochemistry , University of Colorado at Boulder , Boulder , CO , 80309-0215.
J Biomol Struct Dyn. 2000;17 Suppl 1:365-9. doi: 10.1080/07391102.2000.10506642.
Abstract Pf1 filamentous phage represent a simple versatile method for generating partially ordered macromolecules in solution. The phage allow tunable degrees of alignment of macromolecules under a wide range of temperature and solvent conditions. The negatively charged phage are ideal for aligning negatively charged nucleic acids and these phage-nucleic acid solutions are stable indefinitely. We have used Pf1 phage to align various DNA and RNA molecules in solution for measurement of dipolar coupling interactions. These dipolar couplings can be used to improve the local structure of nucleic acids. More importantly they also contain information on the global structure, such as DNA bending, which presently cannot be obtained by standard NMR methods. The principles involved in using Pf1 phage to generate solutions of partially order macromolecules will be discussed. The use of (1)H-(1)H, (1)H-(13)C and (1)H-(15)N dipolar couplings for generating angle constraints for structure refinement of nucleic acids will also be discussed.
摘要 Pf1丝状噬菌体是在溶液中生成部分有序大分子的一种简单通用方法。该噬菌体可在广泛的温度和溶剂条件下使大分子达到可调的排列程度。带负电荷的噬菌体是排列带负电荷核酸的理想选择，并且这些噬菌体 - 核酸溶液可无限期稳定。我们已使用Pf1噬菌体在溶液中排列各种DNA和RNA分子，以测量偶极耦合相互作用。这些偶极耦合可用于改善核酸的局部结构。更重要的是，它们还包含有关整体结构的信息，例如DNA弯曲，目前通过标准核磁共振方法无法获得这些信息。将讨论使用Pf1噬菌体生成部分有序大分子溶液所涉及的原理。还将讨论使用（1）H - （1）H、（1）H - （13）C和（1）H - （15）N偶极耦合为核酸结构细化生成角度约束的情况。
