Exploring modular allostery via interchangeable regulatory domains
Exploring modular allostery via interchangeable regulatory domains
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DOI：
10.1073/pnas.1717621115
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发表时间：
2018-03-20
影响因子：
11.1
通讯作者：
Parker, Emily J.
中科院分区：
文献类型：
--
作者：
Fan, Yifei;Cross, Penelope J.;Parker, Emily J.
大多数蛋白质由两个或两个以上的结构域组成，这些结构域来自有限的蛋白质家族。这些结构域经常通过基因融合事件以各种组合重新排列，从而进化出新的蛋白质功能，包括通过整合调控结构域获得蛋白质变构。3-脱氧达拉比诺-庚糖酸7-磷酸合成酶（DAH7PS）是芳香氨基酸生物合成的第一个酶，具有多种变构机制。dah7ps采用具有共享（β / α）(8)催化结构域的共同结构，该结构域可以连接到act样结构域或通过不同机制运行的choris酸突变酶调节结构域。这些各自的结构域通过控制DAH7PS对配体Tyr或预苯酸盐的响应功能来赋予变构调节。从当代DAH7PS蛋白开始，创造了两个蛋白嵌合体，它们具有互换的调节结构域。这两种工程蛋白都具有催化活性，并具有转换配体特异性和非同源调控域传递的变构机制的新功能变构。这种蛋白质结构域的互换性不仅是设计多结构域蛋白质变构的有效方法，而且是创造新的双功能酶的有效方法。
Most proteins comprise two or more domains from a limited suite of protein families. These domains are often rearranged in various combinations through gene fusion events to evolve new protein functions, including the acquisition of protein allostery through the incorporation of regulatory domains. The enzyme 3-deoxy-Darabino- heptulosonate 7-phosphate synthase (DAH7PS) is the first enzyme of aromatic amino acid biosynthesis and displays a diverse range of allosteric mechanisms. DAH7PSs adopt a common architecture with a shared (beta/alpha)(8) catalytic domain which can be attached to an ACT-like or a chorismate mutase regulatory domain that operates via distinct mechanisms. These respective domains confer allosteric regulation by controlling DAH7PS function in response to ligand Tyr or prephenate. Starting with contemporary DAH7PS proteins, two protein chimeras were created, with interchanged regulatory domains. Both engineered proteins were catalytically active and delivered new functional allostery with switched ligand specificity and allosteric mechanisms delivered by their nonhomologous regulatory domains. This interchangeability of protein domains represents an efficient method not only to engineer allostery in multidomain proteins but to create a new bifunctional enzyme.