Caspary W J, Greene J J, Stempel L M, Ts'o P O
Nucleic Acids Res. 1976 Apr;3(4):847-61. doi: 10.1093/nar/3.4.847.
Homopolyribonucleotides and E. coli DNA wer spin labeled with an iodoacetamide-nitroxide compound. The extent of labeling is highly dependent upon the nature of the base and the secondary structure of the nucleic acid. This spin label-polymer linkage is unstable at high temperatures and in phosphate buffers. In order to determine the effect of changes in the environment of nucleic acids on the esr signals of their attached spin labels, the polynucleotides were subjected to temperature and viscosity perturbations. An increase in temperature (T) affects a linear decrease in the anisotropy factor of the esr signal. The log tau (tau = correlation time) versus (1/T) profile is linear with a positive slope when the spin label is attached to single stranded polynucleotides but exhibits discontinuities at certain critical temperatures when attached to the duplexes poly (As-U) and poly (I-Cs). These critical temperatures are lower than the optical Tm. Logarithmic increase in viscosity was found to produce a linear increase in tau in aqueous sucrose solutions.
同聚核糖核苷酸和大肠杆菌DNA用碘乙酰胺-氮氧化物化合物进行自旋标记。标记程度高度依赖于碱基的性质和核酸的二级结构。这种自旋标记-聚合物连接在高温和磷酸盐缓冲液中不稳定。为了确定核酸环境变化对其连接的自旋标记的电子自旋共振(ESR)信号的影响,对多核苷酸进行了温度和粘度扰动。温度(T)升高会导致ESR信号的各向异性因子呈线性下降。当自旋标记连接到单链多核苷酸时,log τ(τ = 相关时间)与(1/T)的关系曲线是线性的,斜率为正,但当连接到双链多聚(As-U)和多聚(I-Cs)时,在某些临界温度处会出现不连续性。这些临界温度低于光学熔解温度(Tm)。发现在蔗糖水溶液中粘度的对数增加会使τ呈线性增加。