Porschke D, Antosiewicz J
Max-Planck-Institut für biophysikalische Chemie, Göttingen, Federal Republic of Germany.
Biophys J. 1990 Aug;58(2):403-11. doi: 10.1016/S0006-3495(90)82386-9.
The structure of six different tRNA molecules has been analyzed in solution by electrooptical measurements and by bead model simulations. The electric dichroism measured as a function of the field strength shows that tRNA's are associated with substantial permanent dipole moments, which are in the range of 1 x 10(-27) cm(identical to 300 D; before correction for the internal directing field). Rotational diffusion time constants of tRNA molecules in their native state at 2 degrees C show a considerable variation. A particularly large value found for tRNA(Tyr) (50 ns) can be explained by its nine additional nucleotide residues. However, remarkable variations remain for tRNA molecules with the standard number of 76 nucleotide residues (tRNA(Phe) [yeast] 41.6 ns, tRNA(Val) [Escherichia coli] 44.9 ns, tRNA(Glu) [E. coli] 46.8 ns; tRNA(Phe) [E. coli] 48.3 ns). These variations indicate modulations of the tertiary structure, which may be due to a change of the L-hinge angle. Bead models are used to simulate both electric and hydrodynamic parameters of tRNA molecules according to the crystal structure of tRNA(Phe) (yeast). The asymmetric distribution of phosphate charges with respect to the center of diffusion leads, under the assumption of a constant charge reduction to 15% by ion condensation, to a theoretical dipole moment of 7.2 x 10(-28) cm, which is in reasonable agreement with the measurements. The dichroism decay curve calculated for tRNA(Phe) (yeast) is also consistent with the measurements and thus the structure in solution and in the crystal must be very similar in this case. However, our measurements also indicate that the structure of some other tRNA's in solution is different, even in cases with the same number of nucleotide residues.
通过电光测量和珠模型模拟,对六种不同的转运RNA(tRNA)分子在溶液中的结构进行了分析。作为场强函数测量的电二色性表明,tRNA与相当大的永久偶极矩相关,其范围为1×10⁻²⁷厘米(相当于300德拜;在对内部导向场进行校正之前)。2℃时天然状态下tRNA分子的旋转扩散时间常数显示出相当大的变化。在tRNA(Tyr)中发现的特别大的值(50纳秒)可以用其额外的九个核苷酸残基来解释。然而,对于具有76个核苷酸残基标准数量的tRNA分子(酵母tRNA(Phe)41.6纳秒、大肠杆菌tRNA(Val)44.9纳秒、大肠杆菌tRNA(Glu)46.8纳秒;大肠杆菌tRNA(Phe)48.3纳秒),仍存在显著差异。这些差异表明三级结构的调制,这可能是由于L型铰链角的变化。根据酵母tRNA(Phe)的晶体结构,使用珠模型来模拟tRNA分子的电学和流体动力学参数。在离子凝聚使电荷恒定减少到15%的假设下,相对于扩散中心的磷酸盐电荷不对称分布导致理论偶极矩为7.2×10⁻²⁸厘米,这与测量结果合理一致。为酵母tRNA(Phe)计算的二色性衰减曲线也与测量结果一致,因此在这种情况下溶液和晶体中的结构一定非常相似。然而,我们的测量也表明,溶液中一些其他tRNA的结构不同,即使在核苷酸残基数量相同的情况下也是如此。