Department of Chemistry, The Scripps Research Institute, Scripps Florida, Jupiter, FL 33458, USA.
J Am Soc Mass Spectrom. 2013 Jul;24(7):1026-36. doi: 10.1007/s13361-013-0633-x. Epub 2013 May 2.
Screening of bead-based split and pool combinatorial chemistry libraries is a powerful approach to aid the discovery of new chemical compounds able to interact with, and modulate the activities of, protein targets of interest. Split and pool synthesis provides for large and well diversified chemical libraries, in this case comprised of oligomers generated from a well-defined starting set. At the end of the synthesis, each bead in the library displays many copies of a unique oligomer sequence. Because the sequence of the oligomer is not known at the time of screening, methods for decoding of the sequence of each screening "hit" are essential. Here we describe an electron-transfer dissociation (ETD) based tandem mass spectrometry approach for the decoding of mass-encoded split and pool libraries. We demonstrate that the newly described "chiral oligomers of pentenoic amides (COPAs)" yield non-sequence-specific product ions upon collisional activated dissociation; however, complete sequence information can be obtained with ETD. To aid in the decoding of libraries from MS and MS/MS data, we have incorporated (79)Br/(81)Br isotope "tags" to differentiate N- and C-terminal product ions. In addition, we have created "Hit-Find," a software program that allows users to generate libraries in silico. The user can then search all possible members of the chemical library for those that fall within a user-defined mass error.
基于珠粒的分割和池组合化学文库的筛选是一种辅助发现能够与感兴趣的蛋白质靶标相互作用并调节其活性的新化合物的有力方法。分割和池合成提供了大而多样化的化学文库，在这种情况下，文库由从一组明确起始物生成的寡聚物组成。在合成结束时，文库中的每个珠粒都显示出许多独特寡聚物序列的副本。由于在筛选时不知道寡聚物的序列，因此对每个筛选“命中”的序列进行解码的方法至关重要。在这里，我们描述了一种基于电子转移解离（ETD）的串联质谱方法，用于解码质量编码的分割和池文库。我们证明，新描述的“戊烯酰胺的手性寡聚物（COPAs）”在碰撞激活解离时产生非序列特异性产物离子；然而，通过 ETD 可以获得完整的序列信息。为了帮助从 MS 和 MS/MS 数据中解码文库，我们已经引入了（79）Br/（81）Br 同位素“标签”来区分 N-和 C-末端产物离子。此外，我们创建了“Hit-Find”，这是一个软件程序，允许用户在计算机上生成文库。用户可以然后在用户定义的质量误差内搜索化学文库中的所有可能成员。
