Heerschap A, Walters J A, Hilbers C W
Biophys Chem. 1985 Aug;22(3):205-17. doi: 10.1016/0301-4622(85)80044-2.
The thermal unfolding of phenylalanine and initiator tRNA from yeast was investigated over a broad range of solution conditions by differential ultraviolet absorption at 260 nm. Under most conditions, the initiator tRNA exhibits two clearly separated transitions in its differential melting curve which were assigned to unfolding of tertiary and secondary structure elements, respectively. The tertiary transition of this tRNA and the overall transition observed for tRNAPhe do not show a maximum in a curve of Tm values plotted as a function of [Na+]. Such a maximum is usually observed for other nucleic acids at about 1 M Na+. In the presence of 5 mM of the divalent cation Mg2+ (or Ca2+), an overall destabilization of the tRNAs is observed when increasing the sodium concentration. The largest fall in Tm (approximately 15 degrees C) is observed for the tertiary transition of the initiator tRNA. Among various cations tested the following efficiency in the overall stabilization of tRNAPhe is observed: spermine greater than spermidine greater than putrescine greater than Na+ (approximately NH4+). Mg2+ is most efficient at concentrations above 5 mM, but below this concentration spermine and spermidine appear to be more efficient. The same hierarchy in stabilizing power of the polyamines and Na+ is observed for both transitions of the initiator tRNA. However, when compared with Mg2+, the polyamines are far less capable of stabilizing the tertiary structure. In contrast, spermine and spermidine are slightly better than Mg2+ in stabilizing the secondary structure. At increasing concentrations of the polyvalent cations (at fixed [Na+] ) the Tm values of the tRNAs attain a constant value.
通过260nm处的紫外差分吸收,在广泛的溶液条件下研究了来自酵母的苯丙氨酸和起始tRNA的热解折叠。在大多数条件下,起始tRNA在其差分熔解曲线中呈现两个明显分开的转变,分别归因于三级结构和二级结构元件的解折叠。该tRNA的三级转变以及观察到的tRNAPhe的整体转变在作为[Na+]函数绘制的Tm值曲线中没有显示出最大值。对于其他核酸,通常在约1M Na+时观察到这样的最大值。在存在5mM二价阳离子Mg2+(或Ca2+)的情况下,当增加钠浓度时,观察到tRNA的整体去稳定化。对于起始tRNA的三级转变,观察到Tm的最大下降(约15℃)。在测试的各种阳离子中,观察到以下在tRNAPhe整体稳定化方面的效率:精胺大于亚精胺大于腐胺大于Na+(约等于NH4+)。Mg2+在浓度高于5mM时最有效,但低于该浓度时,精胺和亚精胺似乎更有效。对于起始tRNA的两个转变,在多胺和Na+的稳定能力方面观察到相同的等级制度。然而,与Mg2+相比,多胺稳定三级结构的能力要差得多。相反,在稳定二级结构方面,精胺和亚精胺略优于Mg2+。在多价阳离子浓度增加时(在固定的[Na+]下),tRNA的Tm值达到恒定值。