Structural and biophysical characterization of Bacillus thuringiensis insecticidal proteins Cry34Ab1 and Cry35Ab1.
Structural and biophysical characterization of Bacillus thuringiensis insecticidal proteins Cry34Ab1 and Cry35Ab1.
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DOI：
10.1371/journal.pone.0112555
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Narva KE
中科院分区：
文献类型：
--
作者：
Kelker MS;Berry C;Evans SL;Pai R;McCaskill DG;Wang NX;Russell JC;Baker MD;Yang C;Pflugrath JW;Wade M;Wess TJ;Narva KE
众所周知，苏云金芽孢杆菌菌株在孢子形成时产生杀虫蛋白，这些蛋白质沉积在伴孢晶体内含物中。这些昆虫特异性毒素中的大多数在成熟毒素序列中表现出三个结构域。然而，其他Cry毒素在结构上和进化上与这个三结构域家族无关，并且对其三维结构知之甚少，限制了我们对其作用机制的理解以及我们改造蛋白质以增强其功能的能力。在非三结构域Cry毒素中，来自B.苏云金杆菌菌株PS149 B1需要一起作用以通过成孔作用机制对西方玉米根虫（WCR）玉米根萤叶甲产生毒性。Cry 34 Ab 1是一种分子量为144 kDa的蛋白质，具有已知膜破坏活性的细菌溶血素家族（Pfam 06355）的特征，而Cry 35 Ab 1是toxin_10家族（Pfam 05431）的分子量为444 kDa的成员，该家族包括其他杀虫蛋白，例如二元毒素BinA/BinB。Cry 34 Ab 1/Cry 35 Ab 1蛋白代表了一种重要的种子性状技术，已被开发为商业化玉米杂交种中的抗虫性状，用于控制WCR。Cry 34 Ab 1和Cry 35 Ab 1的结构已分别解析到2.15 μ m和1.80 μ m。通过小角X射线散射和自然电喷雾离子迁移质谱进一步研究了毒素的溶液结构。我们在这里提出的第一个公布的结构从aegerolysin蛋白质结构域家族和Cry 34 Ab 1和Cry 35 Ab 1与其他孔形成毒素的结构比较。
Bacillus thuringiensis strains are well known for the production of insecticidal proteins upon sporulation and these proteins are deposited in parasporal crystalline inclusions. The majority of these insect-specific toxins exhibit three domains in the mature toxin sequence. However, other Cry toxins are structurally and evolutionarily unrelated to this three-domain family and little is known of their three dimensional structures, limiting our understanding of their mechanisms of action and our ability to engineer the proteins to enhance their function. Among the non-three domain Cry toxins, the Cry34Ab1 and Cry35Ab1 proteins from B. thuringiensis strain PS149B1 are required to act together to produce toxicity to the western corn rootworm (WCR) Diabrotica virgifera virgifera Le Conte via a pore forming mechanism of action. Cry34Ab1 is a protein of ∼14 kDa with features of the aegerolysin family (Pfam06355) of proteins that have known membrane disrupting activity, while Cry35Ab1 is a ∼44 kDa member of the toxin_10 family (Pfam05431) that includes other insecticidal proteins such as the binary toxin BinA/BinB. The Cry34Ab1/Cry35Ab1 proteins represent an important seed trait technology having been developed as insect resistance traits in commercialized corn hybrids for control of WCR. The structures of Cry34Ab1 and Cry35Ab1 have been elucidated to 2.15 Å and 1.80 Å resolution, respectively. The solution structures of the toxins were further studied by small angle X-ray scattering and native electrospray ion mobility mass spectrometry. We present here the first published structure from the aegerolysin protein domain family and the structural comparisons of Cry34Ab1 and Cry35Ab1 with other pore forming toxins.
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影响因子：
5.7
作者：
Athanasiadis, A;Anderluh, G;Turk, D
通讯作者：
Turk, D
影响因子：
4.2
作者：
Jones RT;Sanchez-Contreras M;Vlisidou I;Amos MR;Yang G;Muñoz-Berbel X;Upadhyay A;Potter UJ;Joyce SA;Ciche TA;Jenkins AT;Bagby S;Ffrench-Constant RH;Waterfield NR
通讯作者：
Waterfield NR
影响因子：
2.8
作者：
Alvarez, Carlos;Mancheno, Jose M.;Lanio, Maria E.
通讯作者：
Lanio, Maria E.
影响因子：
7.4
作者：
Campuzano, Iain;Bush, Matthew F.;Kim, Hugh I.
通讯作者：
Kim, Hugh I.
影响因子：
3.8
作者：
Boonyos, Patcharaporn;Soonsanga, Sumarin;Promdonkoy, Boonhiang
通讯作者：
Promdonkoy, Boonhiang