Flosadóttir Helga Dögg, Stano Michal, Ingólfsson Oddur
Department of Chemistry, University of Iceland, Science Institute, Reykjavík, Iceland.
J Am Soc Mass Spectrom. 2009 Apr;20(4):689-96. doi: 10.1016/j.jasms.2008.12.010. Epub 2008 Dec 24.
Metastable fragmentation of the positively charged, hexameric oligonucleotides 5'-d(TTXYTT) (X and Y are dC, dG, or dA) and 5'-d(CTCGTT), 5'-d(TTCGTC) and 5'-d(CTCGTC) is studied after matrix assisted laser desorption/ionization (MALDI). The influence of the degree of sodiation, i.e., when the acidic protons are one by one exchanged against sodium ions, is systematically studied for the exchange of up to seven protons against sodium ions. Exchanging the acidic protons against sodium gradually quenches the backbone cleavage through the w and a-B channels, and quantitative quenching of these channels is generally achieved with the exchange of four protons against sodium ions. At the same time, the exchange of protons against sodium ions promotes the loss of a neutral, high proton affinity base. The formation of the w and a-B fragments is found to be highly dependent on the sequence of the central bases. A single mechanism consistent with these observations is proposed. In addition to the quenching of the classical w and a-B reaction channels, a drastic and abrupt on/off-switching of new reaction channels is observed as the degree of sodiation successively increases. These channels involve selective loss of the two central bases and the excision of a phosphodiester group and a sugar unit from the center of the oligonucleotides. Synchronously, the two terminal fragments recombine to form a tetramer containing the two terminal nucleosides from each end of the hexamer. Possible mechanism explaining these remarkable channels are discussed.
在基质辅助激光解吸/电离(MALDI)后,研究了带正电荷的六聚体寡核苷酸5'-d(TTXYTT)(X和Y为dC、dG或dA)以及5'-d(CTCGTT)、5'-d(TTCGTC)和5'-d(CTCGTC)的亚稳裂解。系统研究了钠化程度的影响,即当酸性质子逐个被钠离子交换时,最多七个质子与钠离子交换的情况。将酸性质子与钠交换会逐渐抑制通过w和α-B通道的主链裂解,通常在四个质子与钠离子交换时可实现这些通道的定量抑制。同时,质子与钠离子的交换促进了具有高质子亲和力的中性碱基的损失。发现w和α-B片段的形成高度依赖于中央碱基的序列。提出了一种与这些观察结果一致的单一机制。除了经典的w和α-B反应通道的抑制外,随着钠化程度的相继增加,还观察到新反应通道的剧烈且突然的开启/关闭切换。这些通道涉及两个中央碱基的选择性损失以及从寡核苷酸中心切除一个磷酸二酯基团和一个糖单元。同时,两个末端片段重新组合形成一个四聚体,该四聚体包含来自六聚体两端的两个末端核苷。讨论了解释这些显著通道的可能机制。