PLoS Pathogens | |
The Bacterial Defensin Resistance Protein MprF Consists of Separable Domains for Lipid Lysinylation and Antimicrobial Peptide Repulsion | |
Petra Staubitz1  Gabriele Hornig1  Christoph M. Ernst1  Andreas Peschel1  Dirk Kraus2  Nagendra N. Mishra3  Soo-Jin Yang3  Arnold S. Bayer3  Hubert Kalbacher4  | |
[1] Cellular and Molecular Microbiology Division, Interfaculty Institute of Microbiology and Infection Medicine, University of Tübingen, Tübingen, Germany;David Geffen School of Medicine at UCLA, Los Angeles, California, United States of America;Division of Infectious Diseases, Los Angeles Biomedical Research Institute at Harbor–University of California at Los Angeles (UCLA) Medical Center, Torrance, California, United States of America;Medical and Natural Sciences Research Center, University of Tübingen, Tübingen, Germany | |
关键词: Staphylococcus aureus; Protein domains; Lipids; Antimicrobial resistance; Bacterial pathogens; Plasmid construction; Biosynthesis; Antimicrobials; | |
DOI : 10.1371/journal.ppat.1000660 | |
学科分类:生物科学(综合) | |
来源: Public Library of Science | |
【 摘 要 】
Many bacterial pathogens achieve resistance to defensin-like cationic antimicrobial peptides (CAMPs) by the multiple peptide resistance factor (MprF) protein. MprF plays a crucial role in Staphylococcus aureus virulence and it is involved in resistance to the CAMP-like antibiotic daptomycin. MprF is a large membrane protein that modifies the anionic phospholipid phosphatidylglycerol with l-lysine, thereby diminishing the bacterial affinity for CAMPs. Its widespread occurrence recommends MprF as a target for novel antimicrobials, although the mode of action of MprF has remained incompletely understood. We demonstrate that the hydrophilic C-terminal domain and six of the fourteen proposed trans-membrane segments of MprF are sufficient for full-level lysyl-phosphatidylglycerol (Lys-PG) production and that several conserved amino acid positions in MprF are indispensable for Lys-PG production. Notably, Lys-PG production did not lead to efficient CAMP resistance and most of the Lys-PG remained in the inner leaflet of the cytoplasmic membrane when the large N-terminal hydrophobic domain of MprF was absent, indicating a crucial role of this protein part. The N-terminal domain alone did not confer CAMP resistance or repulsion of the cationic test protein cytochrome c. However, when the N-terminal domain was coexpressed with the Lys-PG synthase domain either in one protein or as two separate proteins, full-level CAMP resistance was achieved. Moreover, only coexpression of the two domains led to efficient Lys-PG translocation to the outer leaflet of the membrane and to full-level cytochrome c repulsion, indicating that the N-terminal domain facilitates the flipping of Lys-PG. Thus, MprF represents a new class of lipid-biosynthetic enzymes with two separable functional domains that synthesize Lys-PG and facilitate Lys-PG translocation. Our study unravels crucial details on the molecular basis of an important bacterial immune evasion mechanism and it may help to employ MprF as a target for new anti-virulence drugs.
【 授权许可】
CC BY
【 预 览 】
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