title: Antimicrobial Peptide CMA3 Derived from the CA-MA Hybrid Peptide: Antibacterial and Anti-inflammatory Activities with Low Cytotoxicity and Mechanism of Action in Escherichia coli.
abstract: CA-MA is a hybrid antimicrobial peptide (AMP) derived from two naturally occurring AMPs, cecropin A and magainin 2. CA-MA shows strong antimicrobial activity against Gram-negative and Gram-positive bacteria but also exhibits cytotoxicity toward mammalian cells. Our objective was to identify CA-MA analogues with reduced cytotoxicity by systematic replacement of amino acids with positively charged R groups (His and Lys), aliphatic R groups (Leu), or polar R groups (Glu). Among the CA-MA analogues studied (CMA1 to -6), CMA3 showed the strongest antimicrobial activity, including against drug-resistant Escherichia coli and Pseudomonas aeruginosa strains isolated from hospital patients. CMA3 appeared to act by inducing pore formation (toroidal model) in the bacterial membrane. In cytotoxicity assays, CMA3 showed little cytotoxicity toward human red blood cells (hRBCs) or HaCaT cells. Additionally, no fluorescence was released from small or giant unilamellar vesicles exposed to 60 μM CMA3 for 80 s, whereas fluorescence was released within 35 s upon exposure to CA-MA. CMA3 also exerted strong lipopolysaccharide (LPS)-neutralizing activity in RAW 264.7 cells, and BALB/c mice exposed to LPS after infection by Escherichia coli showed improved survival after administration of one 0.5-mg/kg of body weight or 1-mg/kg dose of CMA3. Finally, in a mouse model of septic shock, CMA3 reduced the levels of proinflammatory factors, including both nitric oxide and white blood cells, and correspondingly reduced lung tissue damage. This study suggests that CMA3 is an antimicrobial/antiendotoxin peptide that could serve as the basis for the development of anti-inflammatory and/or antimicrobial agents with low cytotoxicity.
authors.affiliation: Research Center for Proteinaceous Materials (RCPM), Chosun University, Gwangju, Republic of Korea.
authors.forename: Jong-Kook
authors.initials: JK
authors.lastname: Lee
authors.affiliation: Department of Bioinformatics, Kongju National University, Kongju, Republic of Korea.
authors.forename: Chang Ho
authors.initials: CH
authors.lastname: Seo
authors.affiliation: Department of Physics, Alexandru I. Cuza University, Iasi, Romania.
authors.forename: Tudor
authors.initials: T
authors.lastname: Luchian
authors.affiliation: Research Center for Proteinaceous Materials (RCPM), Chosun University, Gwangju, Republic of Korea Department of Biotechnology and BK21-Plus Research Team for Bioactive Control Technology, Chosun University, Gwangju, Republic of Korea y_k_park@chosun.ac.kr.
authors.forename: Yoonkyung
authors.initials: Y
authors.lastname: Park
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D000900:Anti-Bacterial Agents / Q000138:chemical synthesis / Q000494:pharmacology*; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000138:chemical synthesis / Q000494:pharmacology*; D023181:Antimicrobial Cationic Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D002460:Cell Line; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D004912:Erythrocytes / Q000166:cytology / Q000187:drug effects; D004926:Escherichia coli / Q000187:drug effects* / Q000254:growth & development / Q000648:ultrastructure; D004927:Escherichia coli Infections / Q000188:drug therapy* / Q000382:microbiology / Q000401:mortality; D006801:Humans; D015603:Keratinocytes / Q000166:cytology / Q000187:drug effects; D008070:Lipopolysaccharides; D008264:Macrophages / Q000166:cytology / Q000187:drug effects; D054806:Magainins / Q000737:chemistry; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009569:Nitric Oxide / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D015202:Protein Engineering; D011550:Pseudomonas aeruginosa / Q000187:drug effects* / Q000254:growth & development / Q000648:ultrastructure; D012772:Shock, Septic / Q000139:chemically induced / Q000188:drug therapy* / Q000382:microbiology / Q000401:mortality; D016019:Survival Analysis; D029867:Xenopus Proteins / Q000737:chemistry
chemical_list: D000900:Anti-Bacterial Agents; D000894:Anti-Inflammatory Agents, Non-Steroidal; D023181:Antimicrobial Cationic Peptides; D008070:Lipopolysaccharides; D054806:Magainins; D029867:Xenopus Proteins; C053301:magainin 2 peptide, Xenopus; D009569:Nitric Oxide; C031161:cecropin A