Howard F B, Frazier J, Miles H T
Biochemistry. 1976 Aug 24;15(17):3783-95. doi: 10.1021/bi00662a022.
Poly(2-aminoadenylic acid) forms both double and triple helices with poly(uridylic acid) [poly(U)]. The 2-amino group forms a third hydrogen bond, elevating the 2 leads to 1 transition temperature by 33 degrees C. The third strand, however, has about the same stability as poly(A)-2poly(U), as measured by Tm 3 leads to 2. This selective stabilization of the two-stranded helix results in a much greater resolution of the differnt thermal transitions than that observed in analogous polynucleotide systems. In contrast to other A, U systems 3 leads to 1 and 2 leads to 3 transitions are not observed under any conditions, and the triple helix always undergoes a 3 leads to 2 transition even at very high ionic strength. A 1:1 mixture of poly(2NH2A) and poly(U) exhibits no transient formation of 1:2 complex, unlike similar mixtures of poly(A) with poly(U) and poly(T). This difference is evidently due to a more rapid displacement reaction: [poly(2NH2A) + poly(2NH2A)-2poly(U) leads to 2 poly(2NH2A)-poly(U)] With poly(2NH2A) than with poly(A). We describe a method for establishing the combining ratios of polynucleotide complexes which used a computer to calculate the angles of intersection of mixing curves as explicit and continuous functions of the wavelength. The wavelength dispersions of the angles of intersection determine optimum wavelengths for establishing stoichiometry and can also provide reliable negative evidence that presumably plausible complexes are not formed. Analogous computer procedures have been developed to determine wavelengths which are selective for the formation of both 1:1 and 1:2 complexes. Infrared spectra of the 1:1 and 1:2 complexes resemble those of other A, U homoribopolynucleotide helices in having two and three strong bands, respectively, in the region of carbonyl stretching vibrations. CD spectra of the two complexes are unusual in having negative first extrema of moderate intensity. We attribute these extrema to intrastrand interactions of strong, well-resolved transitions at 278 nm (B2u) of the 2-aminoadenine residues. The CD spectra are correlated with those of other polynucleotide helices.
聚(2-氨基腺苷酸)能与聚(尿苷酸)[聚(U)]形成双链和三链螺旋。2-氨基基团形成第三个氢键,使2→1转变温度升高33℃。然而,通过熔解温度(Tm)测定,第三条链的稳定性与聚(A)-2聚(U)大致相同,即3→2转变的情况。双链螺旋的这种选择性稳定作用导致不同热转变的分辨率比在类似的多核苷酸系统中观察到的要高得多。与其他A、U系统不同,在任何条件下都未观察到3→1和2→3转变,即使在非常高的离子强度下,三链螺旋也总是经历3→2转变。聚(2NH₂A)和聚(U)的1:1混合物不像聚(A)与聚(U)以及聚(T)的类似混合物那样会短暂形成1:2复合物。这种差异显然是由于置换反应更快:[聚(2NH₂A)+聚(2NH₂A)-2聚(U)→2聚(2NH₂A)-聚(U)],聚(2NH₂A)的反应比聚(A)的反应更快。我们描述了一种确定多核苷酸复合物结合比例的方法,该方法使用计算机将混合曲线的交点角度计算为波长的显式连续函数。交点角度的波长色散决定了确定化学计量比的最佳波长,还能提供可靠的负面证据,表明可能看似合理的复合物并未形成。已开发出类似的计算机程序来确定对1:1和1:2复合物形成具有选择性的波长。1:1和1:2复合物的红外光谱与其他A、U同聚核糖多核苷酸螺旋的红外光谱相似,在羰基伸缩振动区域分别有两条和三条强带。这两种复合物的圆二色光谱(CD光谱)不同寻常之处在于具有强度适中的负的第一个极值。我们将这些极值归因于2-氨基腺嘌呤残基在278nm(B2u)处强的、分辨率良好的跃迁的链内相互作用。CD光谱与其他多核苷酸螺旋的光谱相关。