Institute of Organic Chemistry and Center for Molecular Biosciences (CMBI), University of Innsbruck, Innsbruck, Austria.
J Am Soc Mass Spectrom. 2010 Jun;21(6):918-29. doi: 10.1016/j.jasms.2010.02.025. Epub 2010 Mar 9.
We have studied the effect of solution additives on hydrolysis and charge state distribution in ESI MS of RNA. Lower and higher charge state ions can be electrosprayed from solutions containing 25 mM piperidine/25 mM imidazole and 1% vol. triethylamine, respectively, with base-catalyzed hydrolysis rates that are sufficiently slow to perform MS/MS experiments. These lower and higher charge state ions are suitable as precursors for CAD and EDD, respectively. We demonstrate nearly complete sequence coverage for 61 nt RNA dissociated by CAD, and 34 nt RNA dissociated by EDD, and suggest a mechanism for backbone fragmentation in EDD of RNA.
我们研究了溶液添加剂对 RNA 的电喷雾解吸电离质谱（ESI MS）中水解和电荷状态分布的影响。在分别含有 25mM 哌啶/25mM 咪唑和 1%三乙胺的溶液中，可以电喷雾出较低和较高电荷状态的离子，其碱基催化水解速率足够慢，可进行 MS/MS 实验。这些较低和较高电荷状态的离子分别适合作为 CAD 和 EDD 的前体。我们证明了通过 CAD 解离的 61nt RNA 和通过 EDD 解离的 34nt RNA 几乎可以完全测序，并提出了 RNA 在 EDD 中产生骨架断裂的机制。
