Complete alanine scanning of the two-component lantibiotic lacticin 3147: generating a blueprint for rational drug design
Complete alanine scanning of the two-component lantibiotic lacticin 3147: generating a blueprint for rational drug design
复制标题
DOI：
10.1111/j.1365-2958.2006.05398.x
复制
发表时间：
2006-11-01
影响因子：
3.6
通讯作者：
Ross, R. Paul
中科院分区：
文献类型：
--
作者：
Cotter, Paul D.;Deegan, Lucy H.;Ross, R. Paul
羊毛硫抗生素是经后修饰的抗微生物肽，其在纳摩尔浓度下具有活性。一些羊毛硫抗生素已显示通过靶向脂质II（细胞壁生物合成的基本前体）起作用。由于羊毛硫抗生素是核糖体合成的，并且适合于定点诱变，因此它们有可能作为生物模板用于生产具有改进功能的新型肽。然而，如果要采用合理的方法来设计新的lantibiotic，则需要对每个氨基酸（和每个结构域）的作用进行评估。迄今为止，还没有硫醚抗生素经过如此严格的分析。为了解决这个问题，我们已经进行了完全扫描诱变的59个氨基酸的每一个乳酸菌素3147，一个双组分lantibiotic通过协同活性的肽LtnA 1（30个氨基酸）和LtnA 2（29个氨基酸）。所有突变均在天然60 kb质粒pMRC 01中原位进行。许多突变导致可检测的生物活性的消除，并且似乎代表了这些和相关肽内的不变核心。然而，值得注意的是，在59个氨基酸中，至少36个可以改变而不导致活性完全丧失。这些中的许多聚集以在肽内形成可变结构域。在这项研究中产生的信息代表了一个蓝图，这将是至关重要的合理设计的lactic-based抗菌化合物。
Lantibiotics are post-translationally modified antimicrobial peptides which are active at nanomolar concentrations. Some lantibiotics have been shown to function by targeting lipid II, the essential precursor of cell wall biosynthesis. Given that lantibiotics are ribosomally synthesized and amenable to site-directed mutagenesis, they have the potential to serve as biological templates for the production of novel peptides with improved functionalities. However, if a rational approach to novel lantibiotic design is to be adopted, an appreciation of the roles of each individual amino acid (and each domain) is required. To date no lantibiotic has been subjected to such rigorous analysis. To address this issue we have carried out complete scanning mutagenesis of each of the 59 amino acids in lacticin 3147, a two-component lantibiotic which acts through the synergistic activity of the peptides LtnA1 (30 amino acids) and LtnA2 (29 amino acids). All mutations were performed in situ in the native 60kb plasmid, pMRC01. A number of mutations resulted in the elimination of detectable bioactivity and seem to represent an invariable core within these and related peptides. Significantly however, of the 59 amino acids, at least 36 can be changed without resulting in a complete loss of activity. Many of these are clustered to form variable domains within the peptides. The information generated in this study represents a blue-print that will be critical for the rational design of lantibiotic-based antimicrobial compounds.