Biotin-c10-AppCH2ppA is an effective new chemical proteomics probe for diadenosine polyphosphate binding proteins
Biotin-c10-AppCH2ppA is an effective new chemical proteomics probe for diadenosine polyphosphate binding proteins
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DOI：
10.1016/j.bmcl.2014.04.076
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发表时间：
2014-07-01
影响因子：
2.7
通讯作者：
Miller, Andrew D.
中科院分区：
文献类型：
--
作者：
Azhar, M. Ameruddin;Wright, Michael;Miller, Andrew D.
在这里，我们报告的合成，稳定的生物素-c10-AppCH（2）ppA共轭涉及一个不寻常的坎尼扎罗反应步骤的合成。该缀合物用于结合来自大肠杆菌细菌细胞裂解物的预期AP（4）A结合蛋白。结合后，这些蛋白质的身份，然后确定顺利的磁性生物淘选和电喷雾质谱法的过程。蛋白质命中似乎是一组确定的应激蛋白相关靶标。虽然该命中列表可能不是唯一的，并且可能随采样条件和生物体状态的性质而变化，但是命中似乎确实与真正的AP（4）A结合蛋白相对应。因此，这些命中代表了一个健全的基础上，构建新的假设有关的细胞的重要性，AP（4）A的细菌细胞和潜在的生物学意义的AP（4）A蛋白结合相互作用。(C)2014由Elsevier Ltd.出版
Here we report on the synthesis of a synthetic, stable biotin-c10-AppCH(2)ppA conjugate involving an unusual Cannizzaro reaction step. This conjugate is used to bind prospective AP(4)A binding proteins from Escherichia coli bacterial cell lyzates. Following binding, identities of these proteins are then determined smoothly by a process of magnetic bio-panning and electrospray mass spectrometry. Protein hits appear to be a definitive set of stress protein related targets. While this hit list may not be exclusive, and may vary with the nature of sampling conditions and organism status, nevertheless hits do appear to correspond with bona fide AP(4)A-binding proteins. Therefore these hits represent a sound basis on which to construct new hypotheses concerning the cellular importance of AP(4)A to bacterial cells and the potential biological significance of AP(4)A-protein binding interactions. (C) 2014 Published by Elsevier Ltd.