Interaction between Functional Domains of Bacillus thuringiensis Insecticidal Crystal Proteins
Interaction between Functional Domains of Bacillus thuringiensis Insecticidal Crystal Proteins
复制标题
苏云金芽孢杆菌杀虫晶体蛋白功能域之间的相互作用
DOI：
10.1128/aem.65.7.2918-2925.1999
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发表时间：
1999
影响因子：
4.4
通讯作者：
R. Laprade
中科院分区：
文献类型：
--
作者：
C. Rang;V. Vachon;R. D. de Maagd;Mario Villalón;J. Schwartz;D. Bosch;R. Frutos;R. Laprade
摘要通过嵌合蛋白的功能分析，研究了苏云金芽孢杆菌Cry1毒素三个结构域之间的相互作用。通过在Cry1Ab、Cry1Ac、Cry1C和Cry1E之间交换编码结构域I或结构域III的区域来制备杂交基因。通过检测纯化的胰酶激活的嵌合毒素对Sf9细胞存活率和质膜通透性的影响来评价其活性。在亲本毒素中，只有Cry1C对这些细胞具有活性，只有具有Cry1C结构域II的嵌合体具有功能。然而，来自Cry1E的结构域I与来自Cry1C的结构域II和III相结合，导致了一种无活性的毒素，表明来自活性毒素的结构域II是活性所必需的，但不是充分的。结构域I来自Cry1Ab或Cry1Ac的嵌合毒素形成的孔略小于结构域I来自Cry1C的毒素形成的孔。因此，嵌合体形成的孔的性质可能是结构域I和结构域II或III之间相互作用的结果。结构域III似乎调节嵌合毒素的活性：来自Cry1Ab的结构域III与Cry1C的结构域I和II相结合，得到了一种比Cry1C更具活性的蛋白质。
ABSTRACT Interactions among the three structural domains of Bacillus thuringiensis Cry1 toxins were investigated by functional analysis of chimeric proteins. Hybrid genes were prepared by exchanging the regions coding for either domain I or domain III among Cry1Ab, Cry1Ac, Cry1C, and Cry1E. The activity of the purified trypsin-activated chimeric toxins was evaluated by testing their effects on the viability and plasma membrane permeability of Sf9 cells. Among the parental toxins, only Cry1C was active against these cells and only chimeras possessing domain II from Cry1C were functional. Combination of domain I from Cry1E with domains II and III from Cry1C, however, resulted in an inactive toxin, indicating that domain II from an active toxin is necessary, but not sufficient, for activity. Pores formed by chimeric toxins in which domain I was from Cry1Ab or Cry1Ac were slightly smaller than those formed by toxins in which domain I was from Cry1C. The properties of the pores formed by the chimeras are therefore likely to result from an interaction between domain I and domain II or III. Domain III appears to modulate the activity of the chimeric toxins: combination of domain III from Cry1Ab with domains I and II of Cry1C gave a protein which was more strongly active than Cry1C.
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DOI：
10.1073/pnas.93.25.14338
发表时间：
1996-12-10
影响因子：
11.1
作者：
Rajamohan, F;Alzate, O;Dean, DH
通讯作者：
Dean, DH
影响因子：
3.1
作者：
M. K. Lee;T. You;A. Curtiss;D. Dean
通讯作者：
M. K. Lee;T. You;A. Curtiss;D. Dean
DOI：
10.1016/s0021-9258(19)50702-5
发表时间：
1992-02
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
M. K. Lee;R. Milne;A. Z. Ge;Donald H. Dean
通讯作者：
M. K. Lee;R. Milne;A. Z. Ge;Donald H. Dean
DOI：
10.1016/s0021-9258(18)45879-6
发表时间：
1992-02
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Dong Wu;A. Aronson
通讯作者：
Dong Wu;A. Aronson
DOI：
10.1073/pnas.90.19.9041
发表时间：
1993-10-01
影响因子：
11.1
作者：
CHEN, XJ;LEE, MK;DEAN, DH
通讯作者：
DEAN, DH