Patkowski A, Eimer W, Dorfmüller T
Molecular Biophysics Laboratory, A. Mickiewicz University, Poznan, Poland.
Biopolymers. 1990;30(9-10):975-83. doi: 10.1002/bip.360300912.
The collective internal dynamics of transfer RNA(Phe) from brewer's yeast in solution was studied by depolarized dynamic light scattering (DDLS). Within the melting region of tRNA the depolarized spectra consist of two Lorentzian, where the narrow (slow) component describes the overall rotation of the macromolecule. The broad component is attributed to the collective reorientation of the bases within the biopolymer. At high temperature only this relaxation process is observed in the spectrum. The viscosity dependence of the collective internal relaxation process is described by the Stokes-Einstein-Debye equation for rotational diffusion. Estimates of the internal orientational pair correlation factor from the integral depolarized intensities of tRNA(Phe) solutions indicates that the observed dynamics correspond to the collective reorientation of approximately 5 bases. A comparison of the results presented with DDLS studies on the aggregation of the mononucleotide guanosine-5'-monophosphate confirms this result. For a further characterization of the relaxation process we studied the effect of hydrostatic pressure (1-1000 bar) on the depolarized spectra of tRNA. While other spectroscopic methods like nmr, fluorescence polarization anisotropy decay, or ESR give information about the very local motion of a single base within the DNA or RNA, this study shows that by DDLS one can characterize collective internal motions of macromolecules.
通过去偏振动态光散射(DDLS)研究了酿酒酵母中转运RNA(Phe)在溶液中的集体内部动力学。在tRNA的解链区域内,去偏振光谱由两个洛伦兹峰组成,其中窄峰(慢成分)描述了大分子的整体旋转。宽峰归因于生物聚合物内碱基的集体重排。在高温下,光谱中仅观察到这种弛豫过程。集体内部弛豫过程的粘度依赖性由用于旋转扩散的斯托克斯 - 爱因斯坦 - 德拜方程描述。根据tRNA(Phe)溶液的积分去偏振强度对内部取向对相关因子的估计表明,观察到的动力学对应于大约5个碱基的集体重排。将本文的结果与关于单核苷酸5'-磷酸鸟苷聚集的DDLS研究进行比较证实了这一结果。为了进一步表征弛豫过程,我们研究了静水压力(1 - 1000巴)对tRNA去偏振光谱的影响。虽然其他光谱方法如核磁共振、荧光偏振各向异性衰减或电子自旋共振提供了关于DNA或RNA内单个碱基非常局部运动的信息,但本研究表明,通过DDLS可以表征大分子的集体内部运动。