Effects of Mutations in the Phenamacril-Binding Site of Fusarium Myosin-1 on Its Motor Function and Phenamacril Sensitivity
Effects of Mutations in the Phenamacril-Binding Site of Fusarium Myosin-1 on Its Motor Function and Phenamacril Sensitivity
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镰刀菌肌球蛋白-1非那可利结合位点突变对其运动功能和非那可利敏感性的影响
DOI：
10.1021/acsomega.0c02886
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发表时间：
2020-08
期刊：
影响因子：
4.1
通讯作者：
Li Xiang-dong
中科院分区：
文献类型：
--
作者：
Ni Tong;Yuan Min;Ji Huan-Hong;Tang Guangfei;Chen Yun;Ma Zhonghua;Li Xiang-dong
苯那普利是一种镰刀菌特异性杀真菌剂，用于镰刀菌头枯病管理。非那普利的靶点是FgMyo1，禾谷镰刀菌中唯一的I类肌球蛋白。FgMyo1基因的S217L点突变是导致FgMyo1高抗性的主要原因。禾谷胶到苯那普利。最近的结构研究表明，非那普利与FgMyo1马达结构域的50 kDa裂缝结合，形成广泛的相互作用，包括非那普利的氰基和S217的羟基之间的氢键。在这里，我们产生了FgMyo1IQ2，一个截短的FgMyo1由运动域和两个IQ基序与F复合组成。禾谷镰刀菌钙调蛋白在昆虫Sf9细胞中的表达。非那普利有效地抑制FgMyo1IQ2的基础和肌动蛋白激活的ATP酶活性，IC 50在微摩尔范围内。FgMyo1IQ2的S217突变显著增加了非那普利的IC 50。S217 T或S217 L均使非那普利的IC 50增加了100倍，而S217 A仅使IC 50增加了104倍。这些结果表明，S217的羟基在非那普利结合中起着重要但非必需的作用，并且在位置217处的庞大侧链在空间上阻碍非那普利结合。另一方面，S217 P，这可能会改变局部构象的phenamacril结合位点，完全取消phenamacril的抑制。由于非那普利的氰基不与FgMyo1形成可辨别的相互作用，而不是与S217羟基的非必需氢键，因此我们提出非那普利的氰基作为开发新型杀真菌剂的关键修饰位点。
Phenamacril is a Fusarium-specific fungicide used for Fusarium head blight management. The target of phenamacril is FgMyo1, the sole class I myosin in Fusarium graminearum. The point mutation S217L in FgMyo1 is responsible for the high resistance of F. graminearum to phenamacril. Recent structural studies have shown that phenamacril binds to the 50 kDa cleft of the FgMyo1 motor domain, forming extensive interactions, including a hydrogen bond between the cyano group of phenamacril and the hydroxyl group of S217. Here, we produced FgMyo1IQ2, a truncated FgMyo1 composed of the motor domain and two IQ motifs complexed with the F. graminearum calmodulin in insect Sf9 cells. Phenamacril potently inhibited both the basal and the actin-activated ATPase activities of FgMyo1IQ2, with an IC50 in a micromolar range. S217 mutations of FgMyo1IQ2 substantially increased the IC50 of phenamacril. S217T or S217L each increased the IC50 of phenamacril for ∼60-fold, while S217A only increased the IC50 for ∼4-fold. These results indicate that the hydroxyl group of S217 plays an important, but nonessential role in phenamacril binding and that the bulky side chain at the position 217 sterically hinders phenamacril binding. On the other hand, S217P, which might alter the local conformation of the phenamacril-binding site, completely abolished the phenamacril inhibition. Because the cyano group of phenamacril does not form discernible interactions with FgMyo1 other than the nonessential hydrogen bond with the S217 hydroxyl group, we propose the cyano group of phenamacril as a key modification site for the development of novel fungicides.
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全基因组测序表明，myosin-5 的突变导致禾谷镰刀菌对杀菌剂 phenamacril 产生抗性。
影响因子：
4.6
作者：
Zheng Z;Hou Y;Cai Y;Zhang Y;Li Y;Zhou M
通讯作者：
Zhou M
DOI：
10.1073/pnas.161285698
发表时间：
2001-07
影响因子：
11.1
作者：
Xiong Liu;N. Osherov;R. Yamashita;H. Brzeska;E. Korn;G. May
通讯作者：
Xiong Liu;N. Osherov;R. Yamashita;H. Brzeska;E. Korn;G. May
影响因子：
4.9
作者：
J. L. Sørensen;A. Benfield;R. Wollenberg;K. Westphal;R. Wimmer;M. R. Nielsen;K. Nielsen;J. Carere;L. Covarelli;G. Beccari;J. Powell;T. Yamashino;H. Kogler;T. E. Sondergaard;D. Gardiner
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J. L. Sørensen;A. Benfield;R. Wollenberg;K. Westphal;R. Wimmer;M. R. Nielsen;K. Nielsen;J. Carere;L. Covarelli;G. Beccari;J. Powell;T. Yamashino;H. Kogler;T. E. Sondergaard;D. Gardiner
影响因子：
4.7
作者：
R. Wollenberg;S. S. Donau-S.;T. T. Nielsen-T.;J. L. Sørensen;H. Giese;R. Wimmer;T. E. Sondergaard
通讯作者：
R. Wollenberg;S. S. Donau-S.;T. T. Nielsen-T.;J. L. Sørensen;H. Giese;R. Wimmer;T. E. Sondergaard
DOI：
10.1074/jbc.m116.746313
发表时间：
2016-07
期刊：
The Journal of Biological Chemistry
影响因子：
--
作者：
D. Langelaan;Janine Liburd;Yidai Yang;Emily Miller;S. Chitayat;Scott W. Crawley;G. Côté;Steven P. Smith
通讯作者：
D. Langelaan;Janine Liburd;Yidai Yang;Emily Miller;S. Chitayat;Scott W. Crawley;G. Côté;Steven P. Smith