title: Structural determinants of antimicrobial and antiplasmodial activity and selectivity in histidine-rich amphipathic cationic peptides.
abstract: Designed histidine-rich amphipathic cationic peptides, such as LAH4, have enhanced membrane disruption and antibiotic properties when the peptide adopts an alignment parallel to the membrane surface. Although this was previously achieved by lowering the pH, here we have designed a new generation of histidine-rich peptides that adopt a surface alignment at neutral pH. In vitro, this new generation of peptides are powerful antibiotics in terms of the concentrations required for antibiotic activity; the spectrum of target bacteria, fungi, and parasites; and the speed with which they kill. Further modifications to the peptides, including the addition of more hydrophobic residues at the N terminus, the inclusion of a helix-breaking proline residue or using D-amino acids as building blocks, modulated the biophysical properties of the peptides and led to substantial changes in toxicity to human and parasite cells but had only a minimal effect on the antibacterial and antifungal activity. Using a range of biophysical methods, in particular solid-state NMR, we show that the peptides are highly efficient at disrupting the anionic lipid component of model membranes. However, we also show that effective pore formation in such model membranes may be related to, but is not essential for, high antimicrobial activity by cationic amphipathic helical peptides. The information in this study comprises a new layer of detail in the understanding of the action of cationic helical antimicrobial peptides and shows that rational design is capable of producing potentially therapeutic membrane active peptides with properties tailored to their function.
authors.affiliation: Universitá Louis Pasteur/CNRS, UMR7177, Institut de Chimie, 4 Rue Blaise Pascal, F-67070 Strasbourg, France, the UPRES EA-3432 Institut de Bactáriologie, Universitá Louis Pasteur-Hôpitaux Universitaires de Strasbourg, 3 Rue Koeberlá F-67000 Strasbourg, France, the UPRES EA-3950 Institut de Parasitologie et de Pathologie Tropicale de la FacultádeMádecine, Universitá Louis Pasteur, 3 Rue Koeberlá F-67000 Strasbourg, France, INSERM Unitá 575, Physiopathologie du Systéme Nerveux, 5 Rue Blaise Pascal, F-67084 Strasbourg, France, CNRS FRE 3087-Gánáthon, 1 Rue de l'Internationale, F-91002, Evry, France, INSERM Unitá 666 and Service Central de Microscopie Electronique, Universitá Louis Pasteur, 11 Rue Humann, F-67085 Strasbourg, France, and the Department of Plant Molecular Biology, University of Lausanne, Biophore Building, CH-1015 Lausanne, Switzerland. Electronic address: james.mason@kcl.ac.uk.
authors.forename: A James
authors.initials: AJ
authors.lastname: Mason
authors.affiliation: Universitá Louis Pasteur/CNRS, UMR7177, Institut de Chimie, 4 Rue Blaise Pascal, F-67070 Strasbourg, France, the UPRES EA-3432 Institut de Bactáriologie, Universitá Louis Pasteur-Hôpitaux Universitaires de Strasbourg, 3 Rue Koeberlá F-67000 Strasbourg, France, the UPRES EA-3950 Institut de Parasitologie et de Pathologie Tropicale de la FacultádeMádecine, Universitá Louis Pasteur, 3 Rue Koeberlá F-67000 Strasbourg, France, INSERM Unitá 575, Physiopathologie du Systéme Nerveux, 5 Rue Blaise Pascal, F-67084 Strasbourg, France, CNRS FRE 3087-Gánáthon, 1 Rue de l'Internationale, F-91002, Evry, France, INSERM Unitá 666 and Service Central de Microscopie Electronique, Universitá Louis Pasteur, 11 Rue Humann, F-67085 Strasbourg, France, and the Department of Plant Molecular Biology, University of Lausanne, Biophore Building, CH-1015 Lausanne, Switzerland.
authors.forename: Wardi
authors.initials: W
authors.lastname: Moussaoui
authors.affiliation: Universitá Louis Pasteur/CNRS, UMR7177, Institut de Chimie, 4 Rue Blaise Pascal, F-67070 Strasbourg, France, the UPRES EA-3432 Institut de Bactáriologie, Universitá Louis Pasteur-Hôpitaux Universitaires de Strasbourg, 3 Rue Koeberlá F-67000 Strasbourg, France, the UPRES EA-3950 Institut de Parasitologie et de Pathologie Tropicale de la FacultádeMádecine, Universitá Louis Pasteur, 3 Rue Koeberlá F-67000 Strasbourg, France, INSERM Unitá 575, Physiopathologie du Systéme Nerveux, 5 Rue Blaise Pascal, F-67084 Strasbourg, France, CNRS FRE 3087-Gánáthon, 1 Rue de l'Internationale, F-91002, Evry, France, INSERM Unitá 666 and Service Central de Microscopie Electronique, Universitá Louis Pasteur, 11 Rue Humann, F-67085 Strasbourg, France, and the Department of Plant Molecular Biology, University of Lausanne, Biophore Building, CH-1015 Lausanne, Switzerland.
authors.forename: Tamer
authors.initials: T
authors.lastname: Abdelrahman
authors.affiliation: Universitá Louis Pasteur/CNRS, UMR7177, Institut de Chimie, 4 Rue Blaise Pascal, F-67070 Strasbourg, France, the UPRES EA-3432 Institut de Bactáriologie, Universitá Louis Pasteur-Hôpitaux Universitaires de Strasbourg, 3 Rue Koeberlá F-67000 Strasbourg, France, the UPRES EA-3950 Institut de Parasitologie et de Pathologie Tropicale de la FacultádeMádecine, Universitá Louis Pasteur, 3 Rue Koeberlá F-67000 Strasbourg, France, INSERM Unitá 575, Physiopathologie du Systéme Nerveux, 5 Rue Blaise Pascal, F-67084 Strasbourg, France, CNRS FRE 3087-Gánáthon, 1 Rue de l'Internationale, F-91002, Evry, France, INSERM Unitá 666 and Service Central de Microscopie Electronique, Universitá Louis Pasteur, 11 Rue Humann, F-67085 Strasbourg, France, and the Department of Plant Molecular Biology, University of Lausanne, Biophore Building, CH-1015 Lausanne, Switzerland.
authors.forename: Alyae
authors.initials: A
authors.lastname: Boukhari
authors.affiliation: Universitá Louis Pasteur/CNRS, UMR7177, Institut de Chimie, 4 Rue Blaise Pascal, F-67070 Strasbourg, France, the UPRES EA-3432 Institut de Bactáriologie, Universitá Louis Pasteur-Hôpitaux Universitaires de Strasbourg, 3 Rue Koeberlá F-67000 Strasbourg, France, the UPRES EA-3950 Institut de Parasitologie et de Pathologie Tropicale de la FacultádeMádecine, Universitá Louis Pasteur, 3 Rue Koeberlá F-67000 Strasbourg, France, INSERM Unitá 575, Physiopathologie du Systéme Nerveux, 5 Rue Blaise Pascal, F-67084 Strasbourg, France, CNRS FRE 3087-Gánáthon, 1 Rue de l'Internationale, F-91002, Evry, France, INSERM Unitá 666 and Service Central de Microscopie Electronique, Universitá Louis Pasteur, 11 Rue Humann, F-67085 Strasbourg, France, and the Department of Plant Molecular Biology, University of Lausanne, Biophore Building, CH-1015 Lausanne, Switzerland.
authors.forename: Philippe
authors.initials: P
authors.lastname: Bertani
authors.affiliation: Universitá Louis Pasteur/CNRS, UMR7177, Institut de Chimie, 4 Rue Blaise Pascal, F-67070 Strasbourg, France, the UPRES EA-3432 Institut de Bactáriologie, Universitá Louis Pasteur-Hôpitaux Universitaires de Strasbourg, 3 Rue Koeberlá F-67000 Strasbourg, France, the UPRES EA-3950 Institut de Parasitologie et de Pathologie Tropicale de la FacultádeMádecine, Universitá Louis Pasteur, 3 Rue Koeberlá F-67000 Strasbourg, France, INSERM Unitá 575, Physiopathologie du Systéme Nerveux, 5 Rue Blaise Pascal, F-67084 Strasbourg, France, CNRS FRE 3087-Gánáthon, 1 Rue de l'Internationale, F-91002, Evry, France, INSERM Unitá 666 and Service Central de Microscopie Electronique, Universitá Louis Pasteur, 11 Rue Humann, F-67085 Strasbourg, France, and the Department of Plant Molecular Biology, University of Lausanne, Biophore Building, CH-1015 Lausanne, Switzerland.
authors.forename: Arnaud
authors.initials: A
authors.lastname: Marquette
authors.affiliation: Universitá Louis Pasteur/CNRS, UMR7177, Institut de Chimie, 4 Rue Blaise Pascal, F-67070 Strasbourg, France, the UPRES EA-3432 Institut de Bactáriologie, Universitá Louis Pasteur-Hôpitaux Universitaires de Strasbourg, 3 Rue Koeberlá F-67000 Strasbourg, France, the UPRES EA-3950 Institut de Parasitologie et de Pathologie Tropicale de la FacultádeMádecine, Universitá Louis Pasteur, 3 Rue Koeberlá F-67000 Strasbourg, France, INSERM Unitá 575, Physiopathologie du Systéme Nerveux, 5 Rue Blaise Pascal, F-67084 Strasbourg, France, CNRS FRE 3087-Gánáthon, 1 Rue de l'Internationale, F-91002, Evry, France, INSERM Unitá 666 and Service Central de Microscopie Electronique, Universitá Louis Pasteur, 11 Rue Humann, F-67085 Strasbourg, France, and the Department of Plant Molecular Biology, University of Lausanne, Biophore Building, CH-1015 Lausanne, Switzerland.
authors.forename: Peiman
authors.initials: P
authors.lastname: Shooshtarizaheh
authors.affiliation: Universitá Louis Pasteur/CNRS, UMR7177, Institut de Chimie, 4 Rue Blaise Pascal, F-67070 Strasbourg, France, the UPRES EA-3432 Institut de Bactáriologie, Universitá Louis Pasteur-Hôpitaux Universitaires de Strasbourg, 3 Rue Koeberlá F-67000 Strasbourg, France, the UPRES EA-3950 Institut de Parasitologie et de Pathologie Tropicale de la FacultádeMádecine, Universitá Louis Pasteur, 3 Rue Koeberlá F-67000 Strasbourg, France, INSERM Unitá 575, Physiopathologie du Systéme Nerveux, 5 Rue Blaise Pascal, F-67084 Strasbourg, France, CNRS FRE 3087-Gánáthon, 1 Rue de l'Internationale, F-91002, Evry, France, INSERM Unitá 666 and Service Central de Microscopie Electronique, Universitá Louis Pasteur, 11 Rue Humann, F-67085 Strasbourg, France, and the Department of Plant Molecular Biology, University of Lausanne, Biophore Building, CH-1015 Lausanne, Switzerland.
authors.forename: Gilles
authors.initials: G
authors.lastname: Moulay
authors.affiliation: Universitá Louis Pasteur/CNRS, UMR7177, Institut de Chimie, 4 Rue Blaise Pascal, F-67070 Strasbourg, France, the UPRES EA-3432 Institut de Bactáriologie, Universitá Louis Pasteur-Hôpitaux Universitaires de Strasbourg, 3 Rue Koeberlá F-67000 Strasbourg, France, the UPRES EA-3950 Institut de Parasitologie et de Pathologie Tropicale de la FacultádeMádecine, Universitá Louis Pasteur, 3 Rue Koeberlá F-67000 Strasbourg, France, INSERM Unitá 575, Physiopathologie du Systéme Nerveux, 5 Rue Blaise Pascal, F-67084 Strasbourg, France, CNRS FRE 3087-Gánáthon, 1 Rue de l'Internationale, F-91002, Evry, France, INSERM Unitá 666 and Service Central de Microscopie Electronique, Universitá Louis Pasteur, 11 Rue Humann, F-67085 Strasbourg, France, and the Department of Plant Molecular Biology, University of Lausanne, Biophore Building, CH-1015 Lausanne, Switzerland.
authors.forename: Nelly
authors.initials: N
authors.lastname: Boehm
authors.affiliation: Universitá Louis Pasteur/CNRS, UMR7177, Institut de Chimie, 4 Rue Blaise Pascal, F-67070 Strasbourg, France, the UPRES EA-3432 Institut de Bactáriologie, Universitá Louis Pasteur-Hôpitaux Universitaires de Strasbourg, 3 Rue Koeberlá F-67000 Strasbourg, France, the UPRES EA-3950 Institut de Parasitologie et de Pathologie Tropicale de la FacultádeMádecine, Universitá Louis Pasteur, 3 Rue Koeberlá F-67000 Strasbourg, France, INSERM Unitá 575, Physiopathologie du Systéme Nerveux, 5 Rue Blaise Pascal, F-67084 Strasbourg, France, CNRS FRE 3087-Gánáthon, 1 Rue de l'Internationale, F-91002, Evry, France, INSERM Unitá 666 and Service Central de Microscopie Electronique, Universitá Louis Pasteur, 11 Rue Humann, F-67085 Strasbourg, France, and the Department of Plant Molecular Biology, University of Lausanne, Biophore Building, CH-1015 Lausanne, Switzerland.
authors.forename: Bernard
authors.initials: B
authors.lastname: Guerold
authors.affiliation: Universitá Louis Pasteur/CNRS, UMR7177, Institut de Chimie, 4 Rue Blaise Pascal, F-67070 Strasbourg, France, the UPRES EA-3432 Institut de Bactáriologie, Universitá Louis Pasteur-Hôpitaux Universitaires de Strasbourg, 3 Rue Koeberlá F-67000 Strasbourg, France, the UPRES EA-3950 Institut de Parasitologie et de Pathologie Tropicale de la FacultádeMádecine, Universitá Louis Pasteur, 3 Rue Koeberlá F-67000 Strasbourg, France, INSERM Unitá 575, Physiopathologie du Systéme Nerveux, 5 Rue Blaise Pascal, F-67084 Strasbourg, France, CNRS FRE 3087-Gánáthon, 1 Rue de l'Internationale, F-91002, Evry, France, INSERM Unitá 666 and Service Central de Microscopie Electronique, Universitá Louis Pasteur, 11 Rue Humann, F-67085 Strasbourg, France, and the Department of Plant Molecular Biology, University of Lausanne, Biophore Building, CH-1015 Lausanne, Switzerland.
authors.forename: Ruairidh J H
authors.initials: RJH
authors.lastname: Sawers
authors.affiliation: Universitá Louis Pasteur/CNRS, UMR7177, Institut de Chimie, 4 Rue Blaise Pascal, F-67070 Strasbourg, France, the UPRES EA-3432 Institut de Bactáriologie, Universitá Louis Pasteur-Hôpitaux Universitaires de Strasbourg, 3 Rue Koeberlá F-67000 Strasbourg, France, the UPRES EA-3950 Institut de Parasitologie et de Pathologie Tropicale de la FacultádeMádecine, Universitá Louis Pasteur, 3 Rue Koeberlá F-67000 Strasbourg, France, INSERM Unitá 575, Physiopathologie du Systéme Nerveux, 5 Rue Blaise Pascal, F-67084 Strasbourg, France, CNRS FRE 3087-Gánáthon, 1 Rue de l'Internationale, F-91002, Evry, France, INSERM Unitá 666 and Service Central de Microscopie Electronique, Universitá Louis Pasteur, 11 Rue Humann, F-67085 Strasbourg, France, and the Department of Plant Molecular Biology, University of Lausanne, Biophore Building, CH-1015 Lausanne, Switzerland.
authors.forename: Antoine
authors.initials: A
authors.lastname: Kichler
authors.affiliation: Universitá Louis Pasteur/CNRS, UMR7177, Institut de Chimie, 4 Rue Blaise Pascal, F-67070 Strasbourg, France, the UPRES EA-3432 Institut de Bactáriologie, Universitá Louis Pasteur-Hôpitaux Universitaires de Strasbourg, 3 Rue Koeberlá F-67000 Strasbourg, France, the UPRES EA-3950 Institut de Parasitologie et de Pathologie Tropicale de la FacultádeMádecine, Universitá Louis Pasteur, 3 Rue Koeberlá F-67000 Strasbourg, France, INSERM Unitá 575, Physiopathologie du Systéme Nerveux, 5 Rue Blaise Pascal, F-67084 Strasbourg, France, CNRS FRE 3087-Gánáthon, 1 Rue de l'Internationale, F-91002, Evry, France, INSERM Unitá 666 and Service Central de Microscopie Electronique, Universitá Louis Pasteur, 11 Rue Humann, F-67085 Strasbourg, France, and the Department of Plant Molecular Biology, University of Lausanne, Biophore Building, CH-1015 Lausanne, Switzerland.
authors.forename: Marie-Háléne
authors.initials: MH
authors.lastname: Metz-Boutigue
authors.affiliation: Universitá Louis Pasteur/CNRS, UMR7177, Institut de Chimie, 4 Rue Blaise Pascal, F-67070 Strasbourg, France, the UPRES EA-3432 Institut de Bactáriologie, Universitá Louis Pasteur-Hôpitaux Universitaires de Strasbourg, 3 Rue Koeberlá F-67000 Strasbourg, France, the UPRES EA-3950 Institut de Parasitologie et de Pathologie Tropicale de la FacultádeMádecine, Universitá Louis Pasteur, 3 Rue Koeberlá F-67000 Strasbourg, France, INSERM Unitá 575, Physiopathologie du Systéme Nerveux, 5 Rue Blaise Pascal, F-67084 Strasbourg, France, CNRS FRE 3087-Gánáthon, 1 Rue de l'Internationale, F-91002, Evry, France, INSERM Unitá 666 and Service Central de Microscopie Electronique, Universitá Louis Pasteur, 11 Rue Humann, F-67085 Strasbourg, France, and the Department of Plant Molecular Biology, University of Lausanne, Biophore Building, CH-1015 Lausanne, Switzerland.
authors.forename: Ermanno
authors.initials: E
authors.lastname: Candolfi
authors.affiliation: Universitá Louis Pasteur/CNRS, UMR7177, Institut de Chimie, 4 Rue Blaise Pascal, F-67070 Strasbourg, France, the UPRES EA-3432 Institut de Bactáriologie, Universitá Louis Pasteur-Hôpitaux Universitaires de Strasbourg, 3 Rue Koeberlá F-67000 Strasbourg, France, the UPRES EA-3950 Institut de Parasitologie et de Pathologie Tropicale de la FacultádeMádecine, Universitá Louis Pasteur, 3 Rue Koeberlá F-67000 Strasbourg, France, INSERM Unitá 575, Physiopathologie du Systéme Nerveux, 5 Rue Blaise Pascal, F-67084 Strasbourg, France, CNRS FRE 3087-Gánáthon, 1 Rue de l'Internationale, F-91002, Evry, France, INSERM Unitá 666 and Service Central de Microscopie Electronique, Universitá Louis Pasteur, 11 Rue Humann, F-67085 Strasbourg, France, and the Department of Plant Molecular Biology, University of Lausanne, Biophore Building, CH-1015 Lausanne, Switzerland.
authors.forename: Gilles
authors.initials: G
authors.lastname: Právost
authors.affiliation: Universitá Louis Pasteur/CNRS, UMR7177, Institut de Chimie, 4 Rue Blaise Pascal, F-67070 Strasbourg, France, the UPRES EA-3432 Institut de Bactáriologie, Universitá Louis Pasteur-Hôpitaux Universitaires de Strasbourg, 3 Rue Koeberlá F-67000 Strasbourg, France, the UPRES EA-3950 Institut de Parasitologie et de Pathologie Tropicale de la FacultádeMádecine, Universitá Louis Pasteur, 3 Rue Koeberlá F-67000 Strasbourg, France, INSERM Unitá 575, Physiopathologie du Systéme Nerveux, 5 Rue Blaise Pascal, F-67084 Strasbourg, France, CNRS FRE 3087-Gánáthon, 1 Rue de l'Internationale, F-91002, Evry, France, INSERM Unitá 666 and Service Central de Microscopie Electronique, Universitá Louis Pasteur, 11 Rue Humann, F-67085 Strasbourg, France, and the Department of Plant Molecular Biology, University of Lausanne, Biophore Building, CH-1015 Lausanne, Switzerland.
authors.forename: Burkhard
authors.initials: B
authors.lastname: Bechinger
mesh_terms: D000890:Anti-Infective Agents / Q000737:chemistry*; D000962:Antimalarials / Q000737:chemistry*; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry*; D002462:Cell Membrane / Q000737:chemistry; D006639:Histidine / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D057927:Hydrophobic and Hydrophilic Interactions; D010455:Peptides / Q000737:chemistry*; D017433:Protein Structure, Secondary; D013329:Structure-Activity Relationship
chemical_list: D000890:Anti-Infective Agents; D000962:Antimalarials; D023181:Antimicrobial Cationic Peptides; C122154:LAH(4) protein, synthetic; D010455:Peptides; D006639:Histidine