Expanding the diversity of chemical protein modification allows post-translational mimicry
Expanding the diversity of chemical protein modification allows post-translational mimicry
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DOI：
10.1038/nature05757
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发表时间：
2007-04-26
期刊：
影响因子：
64.8
通讯作者：
Davis, Benjamin G.
中科院分区：
文献类型：
--
作者：
van Kasteren, Sander I.;Kramer, Holger B.;Davis, Benjamin G.
当前最重要的科学悖论之一是自然界使用基因的经济性。在所有研究的高等动物中，我们发现的基因比我们先前预期的要少得多。基因最终产物的功能输出似乎比更受限制的蓝图复杂得多。在高等生物中，许多蛋白质的功能都是通过翻译后修饰（PTM）来调节的（1）。氨基酸侧链的这些改变导致更高的结构和功能蛋白质多样性，因此，是解释这种看似不一致的主要竞争者。天然蛋白质生产方法通常产生PTM混合物，其中功能难以剖析或控制。到目前为止，还不可能获得复杂PTM的纯模拟物。在这里，我们报告了一种化学标记的方法，使多个修饰的附着细菌表达（裸）蛋白质支架：这种方法允许重建功能有效的模拟高等生物PTM。通过在广泛使用的LacZ报告酶支架中以适当的距离进行适当的修饰，我们创建了包括用于检测哺乳动物脑炎症和疾病的敏感系统的蛋白质探针。通过靶向合成所需的修饰，化学提供了结构精度和以其他方法不可用的方式用所选PTM重组的能力。通过这种方式，将PTM的化学控制与容易获得的蛋白质支架相结合，为创建蛋白质PTM相互作用的探针提供了系统平台。因此，我们预期构建模型系统2的这种能力将允许解剖一些这种基因产物复杂性，目的是最终能够将特定蛋白质的PTM的模式从体内测定完全复制到体外系统中。
One of the most important current scientific paradoxes is the economy with which nature uses genes. In all higher animals studied, we have found many fewer genes than we would have previously expected. The functional outputs of the eventual products of genes seem to be far more complex than the more restricted blueprint. In higher organisms, the functions of many proteins are modulated by post-translational modifications (PTMs)(1). These alterations of amino-acid side chains lead to higher structural and functional protein diversity and are, therefore, a leading contender for an explanation for this seeming incongruity. Natural protein production methods typically produce PTM mixtures within which function is difficult to dissect or control. Until now it has not been possible to access pure mimics of complex PTMs. Here we report a chemical tagging approach that enables the attachment of multiple modifications to bacterially expressed ( bare) protein scaffolds: this approach allows reconstitution of functionally effective mimics of higher organism PTMs. By attaching appropriate modifications at suitable distances in the widely-used LacZ reporter enzyme scaffold, we created protein probes that included sensitive systems for detection of mammalian brain inflammation and disease. Through target synthesis of the desired modification, chemistry provides a structural precision and an ability to retool with a chosen PTM in a manner not available to other approaches. In this way, combining chemical control of PTM with readily available protein scaffolds provides a systematic platform for creating probes of protein PTM interactions. We therefore anticipate that this ability to build model systems 2 will allow some of this gene product complexity to be dissected, with the aim of eventually being able to completely duplicate the patterns of a particular protein's PTMs from an in vivo assay into an in vitro system.