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Fundamental studies on development of chemotherapeutics targeting cell surface proteins which are essential for growth in Escherichia coli

针对大肠杆菌生长所必需的细胞表面蛋白的化疗药物开发的基础研究

基本信息

批准号：
15380056
负责人：
MATSUYAMA Shin-ichi
金额：
$ 9.47万
依托单位：
Rikkyo University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2006
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-15380056/
关键词：
Escherichia coli cell surface outer membrane inner membrane lipoprotein biosynthesis localization chemotherapentic

项目摘要

Fruitful results described in (1)〜(8) were obtained during the period of this research.(1) The crystal structures of LolA and Lo1B which are involved in the localization of lipoproteins, a major component of the outer membrane, were determined.(2) We showed that the Lol avoidance signal requires a critical length of negative charge at the second residue, and that phosphatidylethanolamine is important for the Lol avoidance function.(3) Five Trp residues which are conserved among Lo1B homologs in Gram-negative bacteria were shown to be important for the receptor activity.(4) We verified the mechanism underlying lipoprotein transfer from LolA to Lo1B is unidirectional and very efficient, but requires no energy input.(5) The random mutagenesis of 14 residues on LolA revealed crucial roles of the hydrophobic cavity and lid in the binding of lipoproteins and their transfer.(6) On the basis of the result that some LolD mutations lowered the ATPase activity of Lo1CDE without affecting that of LolD, the LolD motif was found to be critical for functional interplay with Lo1C / LolE.(7) Using a new screening method which was developed to identify new antibacterial agents targeting the Lol system, we screened for 2 compounds which inhibit the LolA function.(8) We found that a periplasmic protein, YbiS, is a major transpeptidase involved in the covalent linkage between the Lpp and peptidoglycan, using in vitro assay system. The results described above contribute to development of chemotherapeutics targeting cell surface proteins which are essential for growth in Escherichia coli

在本研究期间，获得了（1）和（8）中描述的富有成效的结果。(1)LolA和Lo 1B的晶体结构，这是参与定位的脂蛋白，外膜的主要成分，进行了测定。(2)我们发现，Lol回避信号需要在第二个残基的负电荷的临界长度，磷脂酰乙醇胺是重要的Lol回避功能。(3)在革兰氏阴性菌中Lo 1B同源物中保守的五个Trp残基被证明对受体活性是重要的。(4)我们验证了脂蛋白从LolA转移到Lo 1B的机制是单向的，非常有效，但不需要能量输入。(5)LolA上14个残基的随机突变揭示了疏水腔和疏水盖在脂蛋白结合和转移中的关键作用。(6)基于一些LolD突变降低Lo 1CDE的ATP酶活性而不影响LolD的结果，发现LolD基序对于与Lo 1C/ LolE的功能相互作用是关键的。(7)使用开发用于鉴定靶向Lol系统的新抗菌剂的新筛选方法，我们筛选了2种抑制LolA功能的化合物。(8)我们发现，周质蛋白，YbiS，是一个主要的转肽酶参与Lpp和肽聚糖之间的共价键，使用体外测定系统。上述结果有助于开发靶向大肠杆菌生长所必需的细胞表面蛋白的化疗剂