title: Peptoid-Substituted Hybrid Antimicrobial Peptide Derived from Papiliocin and Magainin 2 with Enhanced Bacterial Selectivity and Anti-inflammatory Activity.
abstract: Antimicrobial peptides (AMPs) are important components of the host innate immune system. Papiliocin is a 37-residue AMP purified from larvae of the swallowtail butterfly Papilio xuthus. Magainin 2 is a 23-residue AMP purified from the skin of the African clawed frog Xenopus laevis. We designed an 18-residue hybrid peptide (PapMA) incorporating N-terminal residues 1-8 of papiliocin and N-terminal residues 4-12 of magainin 2, joined by a proline (Pro) hinge. PapMA showed high antimicrobial activity but was cytotoxic to mammalian cells. To decrease PapMA cytotoxicity, we designed a lysine (Lys) peptoid analogue, PapMA-k, which retained high antimicrobial activity but displayed cytotoxicity lower than that of PapMA. Fluorescent dye leakage experiments and confocal microscopy showed that PapMA targeted bacterial cell membranes whereas PapMA-k penetrated bacterial cell membranes. Nuclear magnetic resonance experiments revealed that PapMA contained an N-terminal α-helix from Lys(3) to Lys(7) and a C-terminal α-helix from Lys(10) to Lys(17), with a Pro(9) hinge between them. PapMA-k also had two α-helical structures in the same region connected with a flexible hinge residue at Nlys(9), which existed in a dynamic equilibrium of cis and trans conformers. Using lipopolysaccharide-stimulated RAW264.7 macrophages, the anti-inflammatory activity of PapMA and PapMA-k was confirmed by inhibition of nitric oxide and inflammatory cytokine production. In addition, treatment with PapMA and PapMA-k decreased the level of ultraviolet irradiation-induced expression of genes encoding matrix metalloproteinase-1 (MMP-1), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) in human keratinocyte HaCaT cells. Thus, PapMA and PapMA-k are potent peptide antibiotics with antimicrobial and anti-inflammatory activity, with PapMA-k displaying enhanced bacterial selectivity.
authors.affiliation: †Department of Bioscience and Biotechnology, Konkuk University, Hwayang-dong, Gwangjin-gu, Seoul 143-701, Korea.
authors.forename: Areum
authors.initials: A
authors.lastname: Shin
authors.affiliation: †Department of Bioscience and Biotechnology, Konkuk University, Hwayang-dong, Gwangjin-gu, Seoul 143-701, Korea.
authors.forename: Eunjung
authors.initials: E
authors.lastname: Lee
authors.affiliation: †Department of Bioscience and Biotechnology, Konkuk University, Hwayang-dong, Gwangjin-gu, Seoul 143-701, Korea.
authors.forename: Dasom
authors.initials: D
authors.lastname: Jeon
authors.affiliation: †Department of Bioscience and Biotechnology, Konkuk University, Hwayang-dong, Gwangjin-gu, Seoul 143-701, Korea.
authors.forename: Young-Guen
authors.initials: YG
authors.lastname: Park
authors.affiliation: ‡Division of Magnetic Resonance, Korea Basic Science Institute, 804-1 Yangchung-ri, Ochang, Chungbuk 363-883, Republic of Korea.
authors.forename: Jeong Kyu
authors.initials: JK
authors.lastname: Bang
authors.affiliation: §Department of Chemistry, Konkuk University, Hwayang-dong, Gwangjin-gu, Seoul 143-701, Korea.
authors.forename: Yong-Sun
authors.initials: YS
authors.lastname: Park
authors.affiliation: ∥Department of Bio-Materials, Graduate School, and Department of Cellular and Molecular Medicine, School of Medicine, Chosun University, Gwangju 501-759, Republic of Korea.
authors.forename: Song Yub
authors.initials: SY
authors.lastname: Shin
authors.affiliation: †Department of Bioscience and Biotechnology, Konkuk University, Hwayang-dong, Gwangjin-gu, Seoul 143-701, Korea.
authors.forename: Yangmee
authors.initials: Y
authors.lastname: Kim
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000494:pharmacology*; D000893:Anti-Inflammatory Agents / Q000737:chemistry / Q000494:pharmacology*; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry / Q000494:pharmacology*; D001419:Bacteria / Q000187:drug effects; D002460:Cell Line; D002462:Cell Membrane / Q000187:drug effects; D015195:Drug Design; D006801:Humans; D015850:Interleukin-6 / Q000276:immunology; D008264:Macrophages / Q000187:drug effects / Q000276:immunology; D054806:Magainins / Q000737:chemistry / Q000494:pharmacology*; D051379:Mice; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D034444:Peptoids / Q000737:chemistry* / Q000494:pharmacology; D017433:Protein Structure, Secondary; D013329:Structure-Activity Relationship; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology; D029867:Xenopus Proteins / Q000737:chemistry / Q000494:pharmacology*
chemical_list: D000900:Anti-Bacterial Agents; D000893:Anti-Inflammatory Agents; D023181:Antimicrobial Cationic Peptides; D015850:Interleukin-6; D054806:Magainins; D034444:Peptoids; D014409:Tumor Necrosis Factor-alpha; D029867:Xenopus Proteins; C551727:papiliocin protein, Papilio xuthus; C053301:magainin 2 peptide, Xenopus