Mechanism of activation of bacterial cellulose synthase by cyclic di-GMP.

Mechanism of activation of bacterial cellulose synthase by cyclic di-GMP.
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DOI:
10.1038/nsmb.2803
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发表时间:
2014-05
影响因子:
16.8
通讯作者:
Zimmer J
Zimmer J
中科院分区:
生物学1区
文献类型:
--
作者:
Morgan JL;McNamara JT;Zimmer J

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细菌信号分子环二鸟苷酸(cyclic-di-GMP)刺激细菌纤维素的合成,细菌纤维素常见于生物膜中。细菌纤维素由纤维素合酶BcsA和BcsB亚基组成的复合物合成并转运穿过内膜。在此我们展示了环二鸟苷酸激活的BcsA - B复合物的晶体结构。这些结构显示环二鸟苷酸通过打破一个盐桥解除了该酶的自身抑制状态,否则这个盐桥会束缚一个保守的门控环,该门控环控制着活性位点的底物进入和底物配位。通过诱变破坏盐桥会产生一种组成型激活的纤维素合酶。此外,环二鸟苷酸激活的BcsA - B复合物包含一个新生的纤维素聚合物,其末端葡萄糖单元位于BcsA活性位点上方的一个新位置,处于催化所需的位置。我们对机制的见解是环二鸟苷酸如何变构调节酶功能的首批实例。
The bacterial signaling molecule cyclic-di-GMP stimulates the synthesis of bacterial cellulose, frequently found in biofilms. Bacterial cellulose is synthesized and translocated across the inner membrane by a complex of the cellulose synthase BcsA and BcsB subunits. Here we present crystal structures of the cyclic-di-GMP-activated BcsA–B complex. The structures reveal that cyclic-di-GMP releases an auto-inhibited state of the enzyme by breaking a salt bridge which otherwise tethers a conserved gating loop that controls access to and substrate coordination at the active site. Disrupting the salt bridge by mutagenesis generates a constitutively active cellulose synthase. Additionally, the cyclic-di-GMP activated BcsA–B complex contains a nascent cellulose polymer whose terminal glucose unit rests at a novel location above BcsA’s active site where it is positioned for catalysis. Our mechanistic insights are the first examples of how cyclic-di-GMP allosterically modulates enzymatic functions.
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