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Dynamics and ordering in a spin-labeled oligonucleotide observed by 220 GHz electron paramagnetic resonance.

机译:通过220 GHz电子顺磁共振观察到的自旋标记寡核苷酸的动力学和有序性。

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摘要

The dynamics of a newly synthesized cytosine spin-label and the spin-labeled pentamer TTC*TT have been observed by high-frequency (220 GHz) electron paramagnetic resonance (EPR) in aqueous solution at ambient temperature using only nanomolar amounts of spin-label. Temperature studies were carried out for both labeled species in buffer containing glycerol. The motion of the spin-labeled monomer could be fitted using a model of fully anisotropic rotation (FAR) over the entire temperature range studied. In the single-stranded pentamer, the high-field spectra are best interpreted using a model of microscopic ordering with macroscopic disorder (MOMD) with the probe in a highly nonpolar environment. The observed local order parameters of 0.60-0.70 suggest a micelle-like structure in which the label is tightly packed with the hydrophobic bases. These preliminary studies illustrate how the excellent orientation selectivity of high-field EPR provides new dynamic information about local base motions in DNA, and also how high-field EPR of spin-labels allows one to discriminate accurately between the effects of local versus global motions in spin-labeled macromolecules.
机译:在室温下仅使用纳摩尔量的自旋标记物通过水溶液中的高频(220 GHz)电子顺磁共振(EPR)观察到新合成的胞嘧啶自旋标记物和自旋标记的五聚体TTC * TT的动力学。在含甘油的缓冲液中对两种标记物进行了温度研究。在整个研究温度范围内,可以使用完全各向异性旋转(FAR)模型拟合自旋标记单体的运动。在单链五聚体中,在高度非极性环境中,使用带有宏观无序的微观排序模型(MOMD)的探针可以最好地解释高场光谱。观察到的0.60-0.70的局部有序参数表明胶束状结构,其中标记物与疏水性碱基紧密堆积。这些初步研究表明,高场EPR的出色定向选择性如何提供有关DNA中局部碱基运动的新动态信息,以及自旋标记的高场EPR如何使人们能够准确地区分局部运动与整体运动的影响。自旋标记的大分子。

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