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首页> 外文期刊>Biochemistry >Molecular motion of spin labeled side chains in #alpha#-helices :analysis by variation of side chain structure
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Molecular motion of spin labeled side chains in #alpha#-helices :analysis by variation of side chain structure

机译：＃alpha＃-螺旋中自旋标记侧链的分子运动：通过侧链结构变化的分析

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Two single cysteine substitution mutants at helix surface sites in T4 lysozyme (D72C and V13 iC) have been modified with a series of nitroxide methanethiosulfonate reagents to investigate the structural and dynamical origins of their electron paramagnetic resonance spectra. The novel reagents include 4-substituted derivatives of either the pyrroline or pyrrolidine series of nitroxides. The spectral line shapes were analyzed as a function of side chain structure and temperature using a simulation method with a single order parameter and diffusion rates about three orthogonal axes as parameters. Taken together, the results provide strong support for an anisotropic motional model of the side chain, which was previously proposed from qualitative features of the spectra and crystal structures of spin labeled T4 lysozyme. Site-specific differences in apparent order parameter are interpreted in terms of backbone dynamics modes with characteristic correlation times in the nanosecond or faster time scale. The saturated 4-substituted pyrrolidine nitroxides are shown to be a suitable template for novel “functionalized” side chains designed to mimic salient features of the native side chains they replace.
机译：T4溶菌酶（D72C和V13 iC）的螺旋表面位点上的两个单个半胱氨酸取代突变体已通过一系列硝基氧化甲烷硫代磺酸盐试剂进行了修饰，以研究其电子顺磁共振光谱的结构和动力学起源。新型试剂包括吡咯啉或吡咯烷系列的氮氧化物的4-取代的衍生物。使用具有单阶参数和围绕三个正交轴的扩散速率作为参数的模拟方法，分析谱线形状作为侧链结构和温度的函数。两者合计，结果为侧链的各向异性运动模型提供了有力的支持，该模型先前是根据自旋标记的T4溶菌酶的光谱和晶体结构的定性特征提出的。表观顺序参数中的特定于站点的差异是根据主链动力学模式解释的，其特征相关时间在纳秒或更短的时间范围内。饱和的4-取代的吡咯烷氮氧化物被证明是适用于新型“功能化”侧链的模板，该侧链旨在模拟它们所取代的天然侧链的显着特征。

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