NKCS, a Mutant of the NK-2 Peptide, Causes Severe Distortions and Perforations in Bacterial, But Not Human Model Lipid Membranes

NKCS is an improved mutant of the bioactive peptide NK-2, which shows strong activity against Escherichia coli and low toxicity towards human cells. The different activity demonstrates the relevance of the physico-chemical nature of the target membrane for the biological effect of this peptide. We studied the effect of this potent antimicrobial peptide on model membranes by activity studies, differential scanning calorimetry, single molecule tracking and tracer efflux experiments. We found that NKCS severely distorted, penetrated and perforated model lipid membranes that resembled bacterial membranes, but not those that were similar to human cell membranes. The interactions of NKCS with phosphatidylethanolamine, which is abundant in bacterial membranes, were especially strong and are probably responsible for its antimicrobial activity.

[1]  R. Vitorino,et al.  Antimicrobial peptides: an alternative for innovative medicines? , 2015, Applied Microbiology and Biotechnology.

[2]  H. Sahl,et al.  Aggregates of nisin with various bactoprenol-containing cell wall precursors differ in size and membrane permeation capacity. , 2013, Biochimica et biophysica acta.

[3]  D. Mercer,et al.  Peptides as the next generation of anti-infectives. , 2013, Future medicinal chemistry.

[4]  R. Gallo,et al.  Antimicrobial peptides: Old Molecules with New Ideas , 2011, The Journal of investigative dermatology.

[5]  U. Kubitscheck,et al.  Cell-penetrating HIV1 TAT peptides can generate pores in model membranes. , 2010, Biophysical journal.

[6]  Sebastian Linser,et al.  Interaction of an antimicrobial peptide with membranes: experiments and simulations with NKCS. , 2010, The journal of physical chemistry. B.

[7]  M. C. Cardoso,et al.  Cell-penetrating HIV1 TAT peptides float on model lipid bilayers. , 2009, Biochemistry.

[8]  A. Blume,et al.  Peptide induced demixing in PG/PE lipid mixtures: a mechanism for the specificity of antimicrobial peptides towards bacterial membranes? , 2009, Biochimica et biophysica acta.

[9]  W. Moerner,et al.  Single-molecule motions of oligoarginine transporter conjugates on the plasma membrane of Chinese hamster ovary cells. , 2008, Journal of the American Chemical Society.

[10]  M. Leippe,et al.  The Antimicrobial Peptide NK-2, the Core Region of Mammalian NK-Lysin, Kills Intraerythrocytic Plasmodium falciparum , 2008, Antimicrobial Agents and Chemotherapy.

[11]  D. Phoenix,et al.  The impact of membrane lipid composition on antimicrobial function of an alpha-helical peptide. , 2008, Chemistry and physics of lipids.

[12]  Angel E Garcia,et al.  Molecular dynamics simulations suggest a mechanism for translocation of the HIV-1 TAT peptide across lipid membranes , 2007, Proceedings of the National Academy of Sciences.

[13]  K. Hahm,et al.  Cell selectivity of an antimicrobial peptide melittin diastereomer with D-amino acid in the leucine zipper sequence. , 2007, Journal of biochemistry and molecular biology.

[14]  T. Goldmann,et al.  Rationale for the Design of Shortened Derivatives of the NK-lysin-derived Antimicrobial Peptide NK-2 with Improved Activity against Gram-negative Pathogens* , 2007, Journal of Biological Chemistry.

[15]  G. Pirri,et al.  Antimicrobial peptides: an overview of a promising class of therapeutics , 2007, Central European Journal of Biology.

[16]  S. Gellman,et al.  Role of membrane lipids in the mechanism of bacterial species selective toxicity by two α/β-antimicrobial peptides , 2006 .

[17]  R. Hancock,et al.  Peptide Antimicrobial Agents , 2006, Clinical Microbiology Reviews.

[18]  R. Hancock,et al.  Cationic host defense (antimicrobial) peptides. , 2006, Current opinion in immunology.

[19]  R. Bakalova,et al.  The NK‐lysin derived peptide NK‐2 preferentially kills cancer cells with increased surface levels of negatively charged phosphatidylserine , 2005, FEBS letters.

[20]  S. Levy.  Antibiotic resistance-the problem intensifies. , 2005, Advanced drug delivery reviews.

[21]  Sebastian Linser,et al.  Structural rearrangement of model membranes by the peptide antibiotic NK-2. , 2005, Biochimica et biophysica acta.

[22]  K. Brogden.  Antimicrobial peptides: pore formers or metabolic inhibitors in bacteria? , 2005, Nature Reviews Microbiology.

[23]  Reto Stöcklin,et al.  Anti‐microbial peptides: from invertebrates to vertebrates , 2004, Immunological reviews.

[24]  Regine Willumeit,et al.  Molecular basis for membrane selectivity of NK-2, a potent peptide antibiotic derived from NK-lysin. , 2003, Biochimica et biophysica acta.

[25]  Michael R. Yeaman,et al.  Mechanisms of Antimicrobial Peptide Action and Resistance , 2003, Pharmacological Reviews.

[26]  M. Leippe,et al.  NK-Lysin and Its Shortened Analog NK-2 Exhibit Potent Activities against Trypanosoma cruzi , 2003, Antimicrobial Agents and Chemotherapy.

[27]  M. Zasloff.  Antimicrobial peptides of multicellular organisms , 2002, Nature.

[28]  A. Tossi,et al.  Amphipathic alpha helical antimicrobial peptides. , 2001, European journal of biochemistry.

[29]  Peter Hinterdorfer,et al.  Single molecule microscopy of biomembranes (Review) , 2000, Molecular membrane biology.

[30]  H. Vogel,et al.  Diversity of antimicrobial peptides and their mechanisms of action. , 1999, Biochimica et biophysica acta.

[31]  M. Dathe,et al.  Structural features of helical antimicrobial peptides: their potential to modulate activity on model membranes and biological cells. , 1999, Biochimica et biophysica acta.

[32]  M. Leippe,et al.  Candidacidal activity of shortened synthetic analogs of amoebapores and NK-lysin , 1999, Medical Microbiology and Immunology.

[33]  O. Schneewind,et al.  Instruments of microbial warfare: bacteriocin synthesis, toxicity and immunity. , 1998, Trends in microbiology.

[34]  I. Nes,et al.  Ribosomally synthesized antimicrobial peptides: their function, structure, biogenesis, and mechanism of action , 1997, Archives of Microbiology.

[35]  R. Hancock,et al.  Peptide antibiotics , 1997, The Lancet.

[36]  Werner Baumgartner,et al.  Characterization of Photophysics and Mobility of Single Molecules in a Fluid Lipid Membrane , 1995 .

[37]  V. Mutt,et al.  NK‐lysin, a novel effector peptide of cytotoxic T and NK cells. Structure and cDNA cloning of the porcine form, induction by interleukin 2, antibacterial and antitumour activity. , 1995, The EMBO journal.

[38]  J. Boggs,et al.  Lipid intermolecular hydrogen bonding: influence on structural organization and membrane function. , 1987, Biochimica et biophysica acta.

[39]  G. Ames.  Lipids of Salmonella typhimurium and Escherichia coli: Structure and Metabolism , 1968, Journal of bacteriology.

[40]  H. Sahl,et al.  Defensin-based anti-infective strategies. , 2014, International journal of medical microbiology : IJMM.

[41]  H. Vogel,et al.  Induction of non-lamellar lipid phases by antimicrobial peptides: a potential link to mode of action. , 2010, Chemistry and physics of lipids.

[42]  R. Willumeit,et al.  Antimicrobial Peptides and Their Interactions with Model Membranes , 2010 .

[43]  D. Phoenix,et al.  The impact of membrane lipid composition on antimicrobial function of an alpha-helical peptide. , 2008, Chemistry and physics of lipids.

[44]  S. Gellman,et al.  Role of membrane lipids in the mechanism of bacterial species selective toxicity by two alpha/beta-antimicrobial peptides. , 2006, Biochimica et biophysica acta.

[45]  Michael D. Abràmoff,et al.  Image processing with ImageJ , 2004 .

[46]  Alessandro Tossi,et al.  Amphipathic, α‐helical antimicrobial peptides , 2000 .