6IST

Crystal structure of a wild type endolysin LysIME-EF1

6IST の概要

• エントリーDOI: 10.2210/pdb6ist/pdb
• 分子名称: Lysin, CALCIUM ION, ... (4 entities in total)
• 機能のキーワード: lysin, hydrolase
• 由来する生物種: Enterococcus phage IMEEF1; 詳細
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 50815.98

構造登録者

Ouyang, S.Y. (登録日: 2018-11-19, 公開日: 2019-11-20, 最終更新日: 2024-03-27)

主引用文献

Zhou, B.,Zhen, X.,Zhou, H.,Zhao, F.,Fan, C.,Perculija, V.,Tong, Y.,Mi, Z.,Ouyang, S.
Structural and functional insights into a novel two-component endolysin encoded by a single gene in Enterococcus faecalis phage.
Plos Pathog., 16:e1008394-e1008394, 2020

PubMed Abstract: Using bacteriophage-derived endolysins as an alternative strategy for fighting drug-resistant bacteria has recently been garnering renewed interest. However, their application is still hindered by their narrow spectra of activity. In our previous work, we demonstrated that the endolysin LysIME-EF1 possesses efficient bactericidal activity against multiple strains of Enterococcus faecalis (E. faecalis). Herein, we observed an 8 kDa fragment and hypothesized that it contributes to LysIME-EF1 lytic activity. To examine our hypothesis, we determined the structure of LysIME-EF1 at 1.75 Å resolution. LysIME-EF1 exhibits a unique architecture in which one full-length LysIME-EF1 forms a tetramer with three additional C-terminal cell-wall binding domains (CBDs) that correspond to the abovementioned 8 kDa fragment. Furthermore, we identified an internal ribosomal binding site (RBS) and alternative start codon within LysIME-EF1 gene, which are demonstrated to be responsible for the translation of the truncated CBD. To elucidate the molecular mechanism for the lytic activity of LysIME-EF1, we combined mutagenesis, lytic activity assays and in vivo animal infection experiments. The results confirmed that the additional LysIME-EF1 CBDs are important for LysIME-EF1 architecture and its lytic activity. To our knowledge, this is the first determined structure of multimeric endolysin encoded by a single gene in E. faecalis phages. As such, it may provide valuable insights into designing potent endolysins against the opportunistic pathogen E. faecalis.

PubMed: 32176738
DOI: 10.1371/journal.ppat.1008394

実験手法

X-RAY DIFFRACTION (1.75 Å)