A Novel Cryptic Clostridial Peptide That Kills Bacteria by a Cell Membrane Permeabilization Mechanism
ABSTRACT This work reports detailed characteristics of the antimicrobial peptide Intestinalin (P30), which is derived from the LysC enzyme of Clostridium intestinale strain URNW. The peptide shows a broader antibacterial spectrum than the parental enzyme, showing potent antimicrobial activity agains...
| Main Authors: | , , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
American Society for Microbiology
2022-10-01
|
| Series: | Microbiology Spectrum |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/spectrum.01657-22 |
| _version_ | 1811329728285507584 |
|---|---|
| author | Monika Szadkowska Michal Olewniczak Anna Kloska Elzbieta Jankowska Malgorzata Kapusta Bartosz Rybak Dariusz Wyrzykowski Wioletta Zmudzinska Artur Gieldon Aleksandra Kocot Anna-Karina Kaczorowska Lukasz Nierzwicki Joanna Makowska Tadeusz Kaczorowski Magdalena Plotka |
| author_facet | Monika Szadkowska Michal Olewniczak Anna Kloska Elzbieta Jankowska Malgorzata Kapusta Bartosz Rybak Dariusz Wyrzykowski Wioletta Zmudzinska Artur Gieldon Aleksandra Kocot Anna-Karina Kaczorowska Lukasz Nierzwicki Joanna Makowska Tadeusz Kaczorowski Magdalena Plotka |
| author_sort | Monika Szadkowska |
| collection | DOAJ |
| description | ABSTRACT This work reports detailed characteristics of the antimicrobial peptide Intestinalin (P30), which is derived from the LysC enzyme of Clostridium intestinale strain URNW. The peptide shows a broader antibacterial spectrum than the parental enzyme, showing potent antimicrobial activity against clinical strains of Gram-positive staphylococci and Gram-negative pathogens and causing between 3.04 ± 0.12 log kill for Pseudomonas aeruginosa PAO1 and 7.10 ± 0.05 log kill for multidrug-resistant Acinetobacter baumannii KPD 581 at a 5 μM concentration. Moreover, Intestinalin (P30) prevents biofilm formation and destroys 24-h and 72-h biofilms formed by Acinetobacter baumannii CRAB KPD 205 (reduction levels of 4.28 and 2.62 log CFU/mL, respectively). The activity of Intestinalin is combined with both no cytotoxicity and little hemolytic effect against mammalian cells. The nuclear magnetic resonance and molecular dynamics (MD) data show a high tendency of Intestinalin to interact with the bacterial phospholipid cell membrane. Although positively charged, Intestinalin resides in the membrane and aggregates into small oligomers. Negatively charged phospholipids stabilize peptide oligomers to form water- and ion-permeable pores, disrupting the integrity of bacterial cell membranes. Experimental data showed that Intestinalin interacts with negatively charged lipoteichoic acid (logK based on isothermal titration calorimetry, 7.45 ± 0.44), causes membrane depolarization, and affects membrane integrity by forming large pores, all of which result in loss of bacterial viability. IMPORTANCE Antibiotic resistance is rising rapidly among pathogenic bacteria, becoming a global public health problem that threatens the effectiveness of therapies for many infectious diseases. In this respect, antimicrobial peptides appear to be an interesting alternative to combat bacterial pathogens. Here, we report the characteristics of an antimicrobial peptide (of 30 amino acids) derived from the clostridial LysC enzyme. The peptide showed killing activity against clinical strains of Gram-positive and Gram-negative pathogens. Experimental data and computational modeling showed that this peptide forms transmembrane pores, directly engaging the negatively charged phospholipids of the bacterial cell membrane. Consequently, dissipation of the electrochemical gradient across cell membranes affects many vital processes, such as ATP synthesis, motility, and transport of nutrients. This kind of dysfunction leads to the loss of bacterial viability. Our firm conviction is that the presented study will be a helpful resource in searching for novel antimicrobial peptides that could have the potential to replace conventional antibiotics. |
| first_indexed | 2024-04-13T15:49:01Z |
| format | Article |
| id | doaj.art-c0dd07784eff4e0ba05fc22289d2e8f6 |
| institution | Directory Open Access Journal |
| issn | 2165-0497 |
| language | English |
| last_indexed | 2024-04-13T15:49:01Z |
| publishDate | 2022-10-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | Microbiology Spectrum |
| spelling | doaj.art-c0dd07784eff4e0ba05fc22289d2e8f62022-12-22T02:40:54ZengAmerican Society for MicrobiologyMicrobiology Spectrum2165-04972022-10-0110510.1128/spectrum.01657-22A Novel Cryptic Clostridial Peptide That Kills Bacteria by a Cell Membrane Permeabilization MechanismMonika Szadkowska0Michal Olewniczak1Anna Kloska2Elzbieta Jankowska3Malgorzata Kapusta4Bartosz Rybak5Dariusz Wyrzykowski6Wioletta Zmudzinska7Artur Gieldon8Aleksandra Kocot9Anna-Karina Kaczorowska10Lukasz Nierzwicki11Joanna Makowska12Tadeusz Kaczorowski13Magdalena Plotka14Laboratory of Extremophiles Biology, Department of Microbiology, Faculty of Biology, University of Gdansk, Gdansk, PolandDepartment of Physical Chemistry, Gdansk University of Technology, Gdansk, PolandDepartment of Medical Biology and Genetics, Faculty of Biology, University of Gdansk, Gdansk, PolandDepartment of Biomedical Chemistry, Faculty of Chemistry, University of Gdansk, Gdansk, PolandDepartment of Plant Cytology and Embryology, Faculty of Biology, University of Gdansk, Gdansk, PolandDepartment of Environmental Toxicology, Faculty of Health Sciences with Institute of Maritime and Tropical Medicine, Medical University of Gdansk, Gdansk, PolandDepartment of General and Inorganic Chemistry, Faculty of Chemistry, University of Gdansk, Gdansk, PolandLaboratory of Biopolymer Structure, Intercollegiate Faculty of Biotechnology, University of Gdansk and Medical University of Gdansk, Gdansk, PolandLaboratory of Simulation of Polymers, Department of Theoretical Chemistry, Faculty of Chemistry, University of Gdansk, Gdansk, PolandLaboratory of Extremophiles Biology, Department of Microbiology, Faculty of Biology, University of Gdansk, Gdansk, PolandCollection of Plasmids and Microorganisms, Faculty of Biology, University of Gdansk, Gdansk, PolandDepartment of Physical Chemistry, Gdansk University of Technology, Gdansk, PolandDepartment of General and Inorganic Chemistry, Faculty of Chemistry, University of Gdansk, Gdansk, PolandLaboratory of Extremophiles Biology, Department of Microbiology, Faculty of Biology, University of Gdansk, Gdansk, PolandLaboratory of Extremophiles Biology, Department of Microbiology, Faculty of Biology, University of Gdansk, Gdansk, PolandABSTRACT This work reports detailed characteristics of the antimicrobial peptide Intestinalin (P30), which is derived from the LysC enzyme of Clostridium intestinale strain URNW. The peptide shows a broader antibacterial spectrum than the parental enzyme, showing potent antimicrobial activity against clinical strains of Gram-positive staphylococci and Gram-negative pathogens and causing between 3.04 ± 0.12 log kill for Pseudomonas aeruginosa PAO1 and 7.10 ± 0.05 log kill for multidrug-resistant Acinetobacter baumannii KPD 581 at a 5 μM concentration. Moreover, Intestinalin (P30) prevents biofilm formation and destroys 24-h and 72-h biofilms formed by Acinetobacter baumannii CRAB KPD 205 (reduction levels of 4.28 and 2.62 log CFU/mL, respectively). The activity of Intestinalin is combined with both no cytotoxicity and little hemolytic effect against mammalian cells. The nuclear magnetic resonance and molecular dynamics (MD) data show a high tendency of Intestinalin to interact with the bacterial phospholipid cell membrane. Although positively charged, Intestinalin resides in the membrane and aggregates into small oligomers. Negatively charged phospholipids stabilize peptide oligomers to form water- and ion-permeable pores, disrupting the integrity of bacterial cell membranes. Experimental data showed that Intestinalin interacts with negatively charged lipoteichoic acid (logK based on isothermal titration calorimetry, 7.45 ± 0.44), causes membrane depolarization, and affects membrane integrity by forming large pores, all of which result in loss of bacterial viability. IMPORTANCE Antibiotic resistance is rising rapidly among pathogenic bacteria, becoming a global public health problem that threatens the effectiveness of therapies for many infectious diseases. In this respect, antimicrobial peptides appear to be an interesting alternative to combat bacterial pathogens. Here, we report the characteristics of an antimicrobial peptide (of 30 amino acids) derived from the clostridial LysC enzyme. The peptide showed killing activity against clinical strains of Gram-positive and Gram-negative pathogens. Experimental data and computational modeling showed that this peptide forms transmembrane pores, directly engaging the negatively charged phospholipids of the bacterial cell membrane. Consequently, dissipation of the electrochemical gradient across cell membranes affects many vital processes, such as ATP synthesis, motility, and transport of nutrients. This kind of dysfunction leads to the loss of bacterial viability. Our firm conviction is that the presented study will be a helpful resource in searching for novel antimicrobial peptides that could have the potential to replace conventional antibiotics.https://journals.asm.org/doi/10.1128/spectrum.01657-22antibacterial peptidesendolysinsmembrane potentialmolecular dynamicstoroidal pore model |
| spellingShingle | Monika Szadkowska Michal Olewniczak Anna Kloska Elzbieta Jankowska Malgorzata Kapusta Bartosz Rybak Dariusz Wyrzykowski Wioletta Zmudzinska Artur Gieldon Aleksandra Kocot Anna-Karina Kaczorowska Lukasz Nierzwicki Joanna Makowska Tadeusz Kaczorowski Magdalena Plotka A Novel Cryptic Clostridial Peptide That Kills Bacteria by a Cell Membrane Permeabilization Mechanism Microbiology Spectrum antibacterial peptides endolysins membrane potential molecular dynamics toroidal pore model |
| title | A Novel Cryptic Clostridial Peptide That Kills Bacteria by a Cell Membrane Permeabilization Mechanism |
| title_full | A Novel Cryptic Clostridial Peptide That Kills Bacteria by a Cell Membrane Permeabilization Mechanism |
| title_fullStr | A Novel Cryptic Clostridial Peptide That Kills Bacteria by a Cell Membrane Permeabilization Mechanism |
| title_full_unstemmed | A Novel Cryptic Clostridial Peptide That Kills Bacteria by a Cell Membrane Permeabilization Mechanism |
| title_short | A Novel Cryptic Clostridial Peptide That Kills Bacteria by a Cell Membrane Permeabilization Mechanism |
| title_sort | novel cryptic clostridial peptide that kills bacteria by a cell membrane permeabilization mechanism |
| topic | antibacterial peptides endolysins membrane potential molecular dynamics toroidal pore model |
| url | https://journals.asm.org/doi/10.1128/spectrum.01657-22 |
| work_keys_str_mv | AT monikaszadkowska anovelcrypticclostridialpeptidethatkillsbacteriabyacellmembranepermeabilizationmechanism AT michalolewniczak anovelcrypticclostridialpeptidethatkillsbacteriabyacellmembranepermeabilizationmechanism AT annakloska anovelcrypticclostridialpeptidethatkillsbacteriabyacellmembranepermeabilizationmechanism AT elzbietajankowska anovelcrypticclostridialpeptidethatkillsbacteriabyacellmembranepermeabilizationmechanism AT malgorzatakapusta anovelcrypticclostridialpeptidethatkillsbacteriabyacellmembranepermeabilizationmechanism AT bartoszrybak anovelcrypticclostridialpeptidethatkillsbacteriabyacellmembranepermeabilizationmechanism AT dariuszwyrzykowski anovelcrypticclostridialpeptidethatkillsbacteriabyacellmembranepermeabilizationmechanism AT wiolettazmudzinska anovelcrypticclostridialpeptidethatkillsbacteriabyacellmembranepermeabilizationmechanism AT arturgieldon anovelcrypticclostridialpeptidethatkillsbacteriabyacellmembranepermeabilizationmechanism AT aleksandrakocot anovelcrypticclostridialpeptidethatkillsbacteriabyacellmembranepermeabilizationmechanism AT annakarinakaczorowska anovelcrypticclostridialpeptidethatkillsbacteriabyacellmembranepermeabilizationmechanism AT lukasznierzwicki anovelcrypticclostridialpeptidethatkillsbacteriabyacellmembranepermeabilizationmechanism AT joannamakowska anovelcrypticclostridialpeptidethatkillsbacteriabyacellmembranepermeabilizationmechanism AT tadeuszkaczorowski anovelcrypticclostridialpeptidethatkillsbacteriabyacellmembranepermeabilizationmechanism AT magdalenaplotka anovelcrypticclostridialpeptidethatkillsbacteriabyacellmembranepermeabilizationmechanism AT monikaszadkowska novelcrypticclostridialpeptidethatkillsbacteriabyacellmembranepermeabilizationmechanism AT michalolewniczak novelcrypticclostridialpeptidethatkillsbacteriabyacellmembranepermeabilizationmechanism AT annakloska novelcrypticclostridialpeptidethatkillsbacteriabyacellmembranepermeabilizationmechanism AT elzbietajankowska novelcrypticclostridialpeptidethatkillsbacteriabyacellmembranepermeabilizationmechanism AT malgorzatakapusta novelcrypticclostridialpeptidethatkillsbacteriabyacellmembranepermeabilizationmechanism AT bartoszrybak novelcrypticclostridialpeptidethatkillsbacteriabyacellmembranepermeabilizationmechanism AT dariuszwyrzykowski novelcrypticclostridialpeptidethatkillsbacteriabyacellmembranepermeabilizationmechanism AT wiolettazmudzinska novelcrypticclostridialpeptidethatkillsbacteriabyacellmembranepermeabilizationmechanism AT arturgieldon novelcrypticclostridialpeptidethatkillsbacteriabyacellmembranepermeabilizationmechanism AT aleksandrakocot novelcrypticclostridialpeptidethatkillsbacteriabyacellmembranepermeabilizationmechanism AT annakarinakaczorowska novelcrypticclostridialpeptidethatkillsbacteriabyacellmembranepermeabilizationmechanism AT lukasznierzwicki novelcrypticclostridialpeptidethatkillsbacteriabyacellmembranepermeabilizationmechanism AT joannamakowska novelcrypticclostridialpeptidethatkillsbacteriabyacellmembranepermeabilizationmechanism AT tadeuszkaczorowski novelcrypticclostridialpeptidethatkillsbacteriabyacellmembranepermeabilizationmechanism AT magdalenaplotka novelcrypticclostridialpeptidethatkillsbacteriabyacellmembranepermeabilizationmechanism |