A bacteriophage endolysin that eliminates intracellular streptococci

eLife

Frank Heinrich¹  Yizhou Yin²  John Moult³  Tarek Vennemann⁴  Daniel C Nelson⁴  Marilia Barros⁴  D Travis Gallagher⁵  Yang Shen⁵  Ryan D Heselpoth⁵  Sara B Linden⁵  Dennis J Spencer⁵  Vincent A Fischetti⁵  Mathias Lösche⁶  David M Donovan⁶ 

[1] Department of Cell Biology and Molecular Genetics, University of Maryland, College Park, Rockville, United States;National Institute of Standards and Technology, Gaithersburg, United States;Animal Biosciences and Biotechnology Lab, Agricultural Research Service, USDA, Beltsville, United States;Department of Physics, Carnegie Mellon University, Pittsburgh, United States;Institute for Bioscience and Biotechnology Research, University of Maryland, College Park, Rockville, United States;Laboratory of Bacterial Pathogenesis and Immunology, The Rockefeller University, New York, United States;

关键词: bacteriophage;    membrane protein;    endolysin;    structure/function;   

DOI  :  10.7554/eLife.13152

来源: DOAJ

【 摘 要 】

PlyC, a bacteriophage-encoded endolysin, lyses Streptococcus pyogenes (Spy) on contact. Here, we demonstrate that PlyC is a potent agent for controlling intracellular Spy that often underlies refractory infections. We show that the PlyC holoenzyme, mediated by its PlyCB subunit, crosses epithelial cell membranes and clears intracellular Spy in a dose-dependent manner. Quantitative studies using model membranes establish that PlyCB interacts strongly with phosphatidylserine (PS), whereas its interaction with other lipids is weak, suggesting specificity for PS as its cellular receptor. Neutron reflection further substantiates that PlyC penetrates bilayers above a PS threshold concentration. Crystallography and docking studies identify key residues that mediate PlyCB–PS interactions, which are validated by site-directed mutagenesis. This is the first report that a native endolysin can traverse epithelial membranes, thus substantiating the potential of PlyC as an antimicrobial for Spy in the extracellular and intracellular milieu and as a scaffold for engineering other functionalities.

【 授权许可】

Unknown