Identification of ribosomal protein L30 as an uncharacterized antimicrobial protein
Identification of ribosomal protein L30 as an uncharacterized antimicrobial protein
复制标题
DOI：
10.1016/j.dci.2021.104067
复制
发表时间：
2021-03-19
影响因子：
2.9
通讯作者：
Ji, Guangdong
中科院分区：
文献类型：
--
作者：
Chen, Ying;Yao, Lan;Ji, Guangdong
几种核糖体蛋白已被证明具有抗菌功能，如抗菌蛋白(AMPs)。然而，这样的信息相当有限，他们的行动模式仍然不明确。在这里，我们证明了文昌鱼RPL30，BjRPL30，是一个未知的AMP，它不仅能通过与内毒素、LTA和PGN相互作用结合革兰氏阴性和革兰氏阳性细菌，而且还能杀死细菌。我们还发现，位于2？46位的残基形成了BjRPL30的抗菌活性核心区。值得注意的是，真核和原核RPL30蛋白中BjRPL302-27和BjRPL3023-46对应的残基的疏水比、净电荷和3D结构都与BjRPL302-27和BjRPL3023-46相似，表明RPL30的抗菌活性是高度保守的。合成的人RPL5-30和RPL26-49也具有抗菌活性的事实进一步证实了这一点。我们证明重组蛋白BjRPL30在体外通过一种膜溶解作用发挥抗菌作用，包括通过内毒素、LTA和PGN与细菌膜相互作用，以及诱导膜去极化。最后，我们发现BjRPL30及其截短型BjRPL302-27和BjRPL3023-46对人红细胞都没有溶血活性，这使它们有可能成为设计新型抗菌AMPS的先导分子。综上所述，RPL30是AMP的一个成员，起源古老，在整个进化过程中高度保守。
Several ribosomal proteins have been shown to adopt for an antimicrobial function as antimicrobial proteins (AMPs). However, information as such is rather limited and their mode of action remains ill-defined. Here we demonstrated that amphioxus RPL30, BjRPL30, was a previously uncharacterized AMP, which was not only capable of binding Gram-negative and Gram-positive bacteria via interaction with LPS, LTA and PGN but also capable of killing the bacteria. We also showed that the residues positioned at 2?46 formed the core region for the antimicrobial activity of BjRPL30. Notably, both the hydrophobic ratio and net charge as well as 3D structures of the residues corresponding to BjRPL302-27 and BjRPL3023-46 from both eukaryotic and prokaryotic RPL30 proteins were closely similar to those of BjRPL302-27 and BjRPL3023-46, suggesting the antibacterial activity of RPL30 was highly conserved. This was further corroborated by the fact that the synthesized counterparts human RPL5-30 and RPL26-49 also had antibacterial activity. We show that the recombinant protein BjRPL30 executes antimicrobial function in vitro by a kind of membranolytic action including interaction with bacterial membrane through LPS, LTA and PGN as well as induction of membrane depolarization. Finally, we found that neither BjRPL30 nor its truncated form BjRPL302-27 and BjRPL3023-46 had hemolytic activity towards human red blood cells, making them promising lead molecules for the design of novel AMPs against bacteria. Altogether, these indicated that RPL30 is a member of AMP which has ancient origin and is highly conserve throughout evolution.