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苏云金芽孢杆菌毒素与小菜蛾敏感和抗性幼虫刷状缘膜囊泡结合的动力学

Kinetics of Bacillus thuringiensis toxin binding with brush border membrane vesicles from susceptible and resistant larvae of Plutella xylostella.

作者信息

Masson L, Mazza A, Brousseau R, Tabashnik B

机构信息

Biotechnology Research Institute, National Research Council of Canada, Montreal, Quebec, Canada.

出版信息

J Biol Chem. 1995 May 19;270(20):11887-96. doi: 10.1074/jbc.270.20.11887.

Abstract

An optical biosensor technology based on surface plasmon resonance was used to determine the kinetic rate constants for interactions between the CryIA(c) toxin from Bacillus thuringiensis and brush border membrane vesicles purified from susceptible and resistant larvae of diamondback moth (Plutella xylostella). CryIA(c) association and dissociation rate constants for vesicles from susceptible larvae were determined to be 4.5 x 10(3) M-1 s-1 and 3.2 x 10(-5) s-1, respectively, resulting in a calculated affinity constant of 7 nM. CryIE toxin did not kill susceptible or resistant larvae and did not bind to brush border vesicles. Contrary to expectations based on previous studies of binding in resistant P. xylostella, the binding kinetics for CryIA(c) did not differ significantly between susceptible larvae and those that were resistant to CryIA(c). Determination of the number of CryIA(c) receptors revealed an approximately 3-fold decrease in total CryIA(c) receptor numbers for resistant vesicles. These results suggest that factors other than binding may be altered in our resistant diamondback moth strain. They also support the view that binding is not sufficient for toxicity.

摘要

基于表面等离子体共振的光学生物传感器技术被用于测定苏云金芽孢杆菌CryIA(c)毒素与从小菜蛾(Plutella xylostella)敏感和抗性幼虫中纯化的刷状缘膜囊泡之间相互作用的动力学速率常数。来自敏感幼虫的囊泡的CryIA(c)结合和解离速率常数分别测定为4.5×10³ M⁻¹ s⁻¹和3.2×10⁻⁵ s⁻¹,计算得出的亲和常数为7 nM。CryIE毒素不能杀死敏感或抗性幼虫,也不与刷状缘囊泡结合。与基于先前对小菜蛾抗性结合研究的预期相反,CryIA(c)在敏感幼虫和对CryIA(c)有抗性的幼虫之间的结合动力学没有显著差异。对CryIA(c)受体数量的测定显示,抗性囊泡的总CryIA(c)受体数量减少了约3倍。这些结果表明,在我们的抗性小菜蛾品系中,除结合之外的其他因素可能发生了改变。它们还支持这样一种观点,即结合不足以产生毒性。

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