Structural characterization and functional analysis of juvenile hormone diol kinase from the silkworm, Bombyx mori.
Structural characterization and functional analysis of juvenile hormone diol kinase from the silkworm, Bombyx mori.
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DOI：
10.1016/j.ijbiomac.2020.11.138
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发表时间：
2020-11
影响因子：
8.2
通讯作者：
Haiyang Xu;Yunshi Zhang;Li Zhang;Zhan Wang;P. Guo;P. Zhao
中科院分区：
文献类型：
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作者：
Haiyang Xu;Yunshi Zhang;Li Zhang;Zhan Wang;P. Guo;P. Zhao
保幼激素二醇激酶(JHDK)是保幼激素代谢途径中的一个重要酶，它催化保幼激素二醇的磷酸化，形成极性代谢物JH二醇磷酸。在此，我们首次报道了家蚕昆虫JHDK的晶体结构，BmJHDK-L2，分辨率为1.22。BmJHDK-L2的结构主要由8个环相连的α-螺旋片段组成，形成4个螺旋-环-螺旋基序。在这四个螺旋-环-螺旋基序中，第一个基序中只有一个钙离子结合。圆二色谱表明BmJHDK-L2具有较强的热稳定性，其热稳定性与二价阳离子无关。BmJHDK-L2的结构进一步允许我们通过计算模拟和结合分析来定义一个ATP结合部位，为JHDK抑制剂的开发提供了结构基础。此外，BmJHDK-L2的表达谱表明，在家蚕的马氏管中保幼激素代谢起主导作用。总而言之，这些发现扩大了我们对昆虫JHDK蛋白的结构和生化特征的了解。
Juvenile hormone diol kinase (JHDK) is an important enzyme involved in the juvenile hormone metabolism pathway, which catalyzes the phosphorylation of juvenile hormone diol to form the polar metabolite JH diol phosphate. Here, we reported the first crystal structure of insect JHDK fromBombyx mori,BmJHDK-L2, determined at a resolution of 1.22 Å. The structure ofBmJHDK-L2 mainly comprises of eight α-helical segments linked with loops, forming four helix-loop-helix motifs. In these four helix-loop-helix motifs with only one calcium ion bound in the first motif. Circular dichroism spectra indicated thatBmJHDK-L2 has strong thermal stability, which is independent of the divalent cation. The structure ofBmJHDK-L2 further allowed us to define an ATP-binding site using computational simulation and binding assays, providing a structural basis for development of inhibitor of JHDK. Moreover, the expression profile ofBmJHDK-L2 indicated a predominant role in juvenile hormone metabolism in the Malpighian tubules of silkworm. Collectively, these findings expand our knowledge regarding the structural and biochemical features of insect JHDK proteins.