Investigating interactions of the Bacillus subtilis spore coat proteins CotY and CotZ using single molecule force spectroscopy
Investigating interactions of the Bacillus subtilis spore coat proteins CotY and CotZ using single molecule force spectroscopy
复制标题
使用单分子力谱研究枯草芽孢杆菌孢子外壳蛋白 CotY 和 CotZ 的相互作用
DOI：
10.1016/j.jsb.2015.09.001
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发表时间：
2015-10-01
影响因子：
3
通讯作者：
Tang, Jilin
中科院分区：
文献类型：
--
作者：
Liu, Huiqing;Krajcikova, Daniela;Tang, Jilin
枯草芽孢杆菌形成的孢子被一层保护性的多层外壳包围，称为被毛，这使孢子能够抵抗各种环境压力并促进孢子萌发。孢子外壳由70多种不同的蛋白质组成，排列成至少四个不同的结构层：下被毛、内被毛、外被毛和外壳。然而，这些蛋白，特别是形态发生蛋白，如何相互作用以建立有组织的、功能性的被膜层仍然知之甚少。CotY和CotZ作为地壳的组成部分，在孢子周围的地壳组装中起着形态形成的作用。本研究采用单分子力谱法在单分子水平上研究了CotY和CotZ之间的相互作用和动力学。结果表明，CotY与CotZ之间存在同型相互作用，CotY与CotZ之间存在异型相互作用。此外，通过监测不同拉速下解结合力与加载速率之间的关系，研究了配合物的解离动力学。通过这种方法，得到了三种不同配合物的一系列动力学参数。CotY与CotZ、CotY与CotY之间存在较强的相互作用，CotZ与CotZ之间存在较弱的相互作用。(C) 2015爱思唯尔公司版权所有。
Spores formed by Bacillus subtilis are surrounded by a protective and multilayered shell, termed the coat, which grants the spores resistance to various environmental stresses and facilitates spore germination. The spore coat consists of more than seventy different proteins, arranged into at least four distinct structural layers: the undercoat, inner coat, outer coat and crust. However, how these proteins, especially the morphogenetic proteins, interact to establish the organized, functional coat layers remains poorly understood. CotY and CotZ as the components of the crust, play a morphogenetic role in the crust assembly around the spore. In this study, the single molecule force spectroscopy was used to investigate the interaction and dynamics between CotY and CotZ at the single-molecule level. The results show that homotypic interactions of CotY and CotZ and the heterotypic interaction between CotY and CotZ exist. Furthermore, the dissociation kinetics of the complexes were studied by monitoring the relationship between the unbinding forces and the loading rates at different pulling velocities. In this way, a series of kinetic parameters regarding the three different complexes were obtained. It revealed the strong interactions between CotY and CotZ, CotY and CotY, and a relatively weak interaction of CotZ and CotZ. (C) 2015 Elsevier Inc. All rights reserved.