Hansen M R, Mueller L, Pardi A
Department of Chemistry and Biochemistry, University of Colorado, Boulder 80309-0215, USA.
Nat Struct Biol. 1998 Dec;5(12):1065-74. doi: 10.1038/4176.
Dipolar coupling interactions represent an extremely valuable source of long-range distance and angle information that was previously not available for solution structure determinations of macromolecules. This is because observation of these dipolar coupling data requires creating an anisotropic environment for the macromolecule. Here we introduce a new method for generating tunable degrees of alignment of macromolecules by addition of magnetically aligned Pf1 filamentous bacteriophage as a cosolute. This phage-induced alignment technique has been used to study 1H-1H, 1H-13C, and 1H-15N dipolar coupling interactions in a DNA duplex, an RNA hairpin and several proteins including thioredoxin and apo-calmodulin. The phage allow alignment of macromolecules over a wide range of temperature and solution conditions and thus represent a stable versatile method for generating partially aligned macromolecules in solution.
偶极耦合相互作用是长程距离和角度信息的极其宝贵的来源，而此前在大分子溶液结构测定中无法获得此类信息。这是因为观察这些偶极耦合数据需要为大分子创造一个各向异性环境。在此，我们引入了一种新方法，即通过添加磁取向的Pf1丝状噬菌体作为共溶质来生成大分子的可调谐排列程度。这种噬菌体诱导排列技术已被用于研究DNA双链体、RNA发夹以及包括硫氧还蛋白和脱钙调蛋白在内的几种蛋白质中的1H-1H、1H-13C和1H-15N偶极耦合相互作用。该噬菌体能够在很宽的温度和溶液条件范围内实现大分子的排列，因此是一种在溶液中生成部分排列大分子的稳定通用方法。
