Specificity, structure and function of the lantibiotic immunity proteins SpaI and NisI

羊毛硫抗生素免疫蛋白 SpaI 和 NisI 的特异性、结构和功能

• 批准号: 240920551
• 负责人: Professor Dr. Karl-Dieter Entian
• 金额: --
• 依托单位: Institut für Molekulare Biowissenschaften
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2013
• 资助国家: 德国
• 起止时间: 2012-12-31 至 2016-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/240920551?language=en
• 关键词: Specificity; structure; function; lantibiotic; immunity

Lantibiotics are peptide antibiotics containing characteristic lanthionine ring structures. They are ribosomally synthesized and post-translationally modified. The antibiotic mode of action of lantibiotics subtilin from Bacillus subtilis and ninin from Lactococcus lactis are based on the binding to the cell wall precursor lipid II and the formation of multimere lantibiotic/lipid II membrane complexes. Lantibiotic producers are sensitive against their own lantibiotic and therefore they express a membrane bound LanI immunity protein and an independent ABC-transporter complex. The mode of action and the molecular mechanism of LanI mediated immunity are still unknown. Although subtilin and nisin have a similar molecular structure, the respective immunity proteins NisI and SpaI are highly selective for the respective antibiotic. We could resolve the NMR-structure of SpaI in solution, with a so far unknown folding pattern. No conclusions on the immunity mechanism could be drawn form the structure so far. At present we consider four possible mechanisms for SpaI mediated immunity, which we want to verify during the project: (A) LanI proteins capture lantibiotics before they reach the cytoplasma membrane (capture function), (B) LanI proteins interact with lantibiotic/lipid II pores to prevent pore formation (complex inhibition), (C) LanI interact and destroy existing lantibiotic/lipid II pores (pore destruction), and (D) SpaI protein seal existing lantibiotic/lipid II pores (pore sealing). Aim of the project is to understand the molecular mechanisms, the high specificity and the membrane interaction of SpaI and NisI proteins by comparing the functional homologue and structural most likely different immunity proteins, using molecular biological, biochemical and biopyhysical methods. Understanding the molecular mechanism and the specificity of SpaI immunity is also important for a possible application of nisin and subtilin variants with altered antibiotic activities.

羊毛硫抗生素是含有特征性羊毛硫氨酸环结构的肽抗生素。它们是核糖体合成的并经过翻译后修饰。来自枯草芽孢杆菌的羊毛硫抗生素枯草菌素和来自乳酸乳球菌的尼宁的抗生素作用模式基于与细胞壁前体脂质 II 的结合以及多聚羊毛硫抗生素/脂质 II 膜复合物的形成。羊毛硫抗生素生产者对其自身的羊毛硫抗生素敏感，因此它们表达膜结合的 LanI 免疫蛋白和独立的 ABC 转运蛋白复合物。 LanI介导的免疫的作用方式和分子机制仍不清楚。尽管枯草菌素和乳酸链球菌素具有相似的分子结构，但各自的免疫蛋白NisI和SpaI对各自的抗生素具有高度选择性。我们可以解析溶液中 SpaI 的 NMR 结构，具有迄今为止未知的折叠模式。迄今为止，还无法从该结构中得出有关免疫机制的结论。目前，我们考虑 SpaI 介导的免疫的四种可能机制，我们希望在项目期间验证这些机制：(A) LanI 蛋白在羊毛硫抗生素到达细胞质膜之前捕获羊毛硫抗生素（捕获功能），(B) LanI 蛋白与羊毛硫抗生素/脂质 II 相互作用孔以防止孔形成（复合抑制），(C) LanI 相互作用并破坏现有的羊毛硫抗生素/脂质 II 孔（孔破坏），以及 (D) SpaI 蛋白密封现有的羊毛硫抗生素/脂质 II 孔（孔密封）。该项目的目的是通过使用分子生物学、生物化学和生物物理方法比较功能同源物和结构最可能不同的免疫蛋白，了解 SpaI 和 NisI 蛋白的分子机制、高特异性和膜相互作用。了解 SpaI 免疫的分子机制和特异性对于改变抗生素活性的乳链菌肽和枯草菌素变体的可能应用也很重要。

项目成果

Specificity of Subtilin-Mediated Activation of Histidine Kinase SpaK

• DOI: 10.1128/aem.00781-17
• 发表时间: 2017-07
• 期刊: Applied and Environmental Microbiology
• 影响因子: 4.4
• 作者: Christoph Geiger;T. Spieß;S. Korn;P. Kötter;K. Entian

NMR resonance assignments of the lantibiotic immunity protein NisI from Lactococcuslactis

乳酸乳球菌羊毛硫抗生素免疫蛋白 NisI 的 NMR 共振归属

• DOI: 10.1007/s12104-015-9595-1
• 发表时间: 2015
• 期刊: Biomolecular NMR Assignments
• 影响因子: 0.9
• 作者: C. Hacker;N. A. Christ;E. Duchardt-Ferner;S. Korn;L. Berninger;P. Kötter;K.-D. Entian;J. Wöhnert
• 通讯作者: J. Wöhnert

Autoinduction Specificities of the Lantibiotics Subtilin and Nisin

羊毛硫抗生素枯草菌素和乳链菌肽的自诱导特性

• DOI: 10.1128/aem.02392-15
• 发表时间: 2015
• 期刊: Applied and Environmental Microbiology
• 影响因子: 4.4
• 作者: T. Spieß;S. M. Korn;P. Kötter;K.-D. Entian
• 通讯作者: K.-D. Entian