Critical amino acids for the insecticidal activity of Vip3Af from Bacillus thuringiensis: Inference on structural aspects.
Critical amino acids for the insecticidal activity of Vip3Af from Bacillus thuringiensis: Inference on structural aspects.
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DOI：
10.1038/s41598-018-25346-3
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发表时间：
2018-05-15
影响因子：
4.6
通讯作者：
Ferré J
中科院分区：
文献类型：
--
作者：
Banyuls N;Hernández-Rodríguez CS;Van Rie J;Ferré J
苏云金芽孢杆菌Vip 3营养期杀虫蛋白是害虫综合防治中的重要工具。虽然对它们的一般作用模式有广泛的共识，但它们的作用模式的细节尚未完全阐明，它们的结构仍然未知。在这项工作中，丙氨酸扫描技术进行了558的Vip 3Af 1蛋白质的788个氨基酸的总数。在558个氨基酸残基中，有19个氨基酸残基被替换，其中19个氨基酸残基被替换后蛋白质表达受损，另外19个氨基酸残基被替换后对草地贪夜蛾的杀虫活性严重受损。后19个取代主要集中在蛋白质序列的两个区域（氨基酸167-272和氨基酸689-741）。大多数取代也降低了对小地老虎的活性。通过SDS-PAGE评估，显示出降低的杀虫活性的突变蛋白对蛋白酶的敏感性的表征揭示了6种不同的带型。对大多数选择的突变体的固有荧光的研究显示，发射峰仅轻微偏移，可能表明三级结构仅发生微小变化。首次提出了Vip 3Af 1的计算机模拟三维结构。
Vip3 vegetative insecticidal proteins from Bacillus thuringiensis are an important tool for crop protection against caterpillar pests in IPM strategies. While there is wide consensus on their general mode of action, the details of their mode of action are not completely elucidated and their structure remains unknown. In this work the alanine scanning technique was performed on 558 out of the total of 788 amino acids of the Vip3Af1 protein. From the 558 residue substitutions, 19 impaired protein expression and other 19 substitutions severely compromised the insecticidal activity against Spodoptera frugiperda. The latter 19 substitutions mainly clustered in two regions of the protein sequence (amino acids 167–272 and amino acids 689–741). Most of these substitutions also decreased the activity to Agrotis segetum. The characterisation of the sensitivity to proteases of the mutant proteins displaying decreased insecticidal activity revealed 6 different band patterns as evaluated by SDS-PAGE. The study of the intrinsic fluorescence of most selected mutants revealed only slight shifts in the emission peak, likely indicating only minor changes in the tertiary structure. An in silico modelled 3D structure of Vip3Af1 is proposed for the first time.
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影响因子：
2.1
作者：
Bhalla, R;Dalal, M;Kumar, PA
通讯作者：
Kumar, PA
影响因子：
2.8
作者：
Calvete, JJ;Marcinkiewicz, C;Sanz, L
通讯作者：
Sanz, L
影响因子：
14.9
作者：
Ferre, F.;Clote, P.
通讯作者：
Clote, P.
影响因子：
14.9
作者：
Ceroni, Alessio;Passerini, Andrea;Vullo, Alessandro;Frasconi, Paolo
通讯作者：
Frasconi, Paolo
影响因子：
4.1
作者：
Calvete, JJ;Moreno-Murciano, MP;Marcinkiewicz, C
通讯作者：
Marcinkiewicz, C