title: Anti-microbial, anti-biofilm activities and cell selectivity of the NRC-16 peptide derived from witch flounder, Glyptocephalus cynoglossus.
abstract: Previous studies had identified novel antimicrobial peptides derived from witch flounder. In this work, we extended the search for the activity of peptide that showed antibacterial activity on clinically isolated bacterial cells and bacterial biofilm. Pseudomonas aeruginosa was obtained from otitis media and cholelithiasis patients, while Staphylococcus aureus was isolated from otitis media patients. We found that synthetic peptide NRC-16 displays antimicrobial activity and is not sensitive to salt during its bactericidal activity. Interestingly, this peptide also led to significant inhibition of biofilm formation at a concentration of 4-16 μM. NRC-16 peptide is able to block biofilm formation at concentrations just above its minimum inhibitory concentration while conventional antibiotics did not inhibit the biofilm formation except ciprofloxacin and piperacillin. It did not cause significant lysis of human RBC, and is not cytotoxic to HaCaT cells and RAW264.7 cells, thereby indicating its selective antimicrobial activity. In addition, the peptide's binding and permeation activities were assessed by tryptophan fluorescence, calcein leakage and circular dichroism using model mammalian membranes composed of phosphatidylcholine (PC), PC/cholesterol (CH) and PC/sphingomyelin (SM). These experiments confirmed that NRC-16 does not interact with any of the liposomes but the control peptide melittin did. Taken together, we found that NRC-16 has potent antimicrobial and antibiofilm activities with less cytotoxicity, and thus can be considered for treatment of microbial infection in the future.
authors.affiliation: Research Center for Proteineous Materials, Chosun University, Gwangju 501-759, Korea.
authors.forename: Ramamourthy
authors.initials: R
authors.lastname: Gopal
authors.forename: Jun Ho
authors.initials: JH
authors.lastname: Lee
authors.forename: Young Gwon
authors.initials: YG
authors.lastname: Kim
authors.forename: Myeong-Sun
authors.initials: MS
authors.lastname: Kim
authors.forename: Chang Ho
authors.initials: CH
authors.lastname: Seo
authors.forename: Yoonkyung
authors.initials: Y
authors.lastname: Park
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000138:chemical synthesis / Q000302:isolation & purification / Q000494:pharmacology*; D023181:Antimicrobial Cationic Peptides / Q000138:chemical synthesis / Q000302:isolation & purification / Q000494:pharmacology*; D018441:Biofilms / Q000187:drug effects*; D002460:Cell Line; D002942:Circular Dichroism / Q000379:methods; D004305:Dose-Response Relationship, Drug; D029941:Fish Proteins / Q000138:chemical synthesis / Q000302:isolation & purification / Q000494:pharmacology*; D005432:Flounder / Q000378:metabolism*; D005452:Fluoresceins / Q000378:metabolism; D005453:Fluorescence; D006461:Hemolysis / Q000187:drug effects; D006801:Humans; D051379:Mice; D008826:Microbial Sensitivity Tests; D010033:Otitis Media / Q000188:drug therapy / Q000382:microbiology; D011550:Pseudomonas aeruginosa / Q000187:drug effects / Q000302:isolation & purification; D013211:Staphylococcus aureus / Q000187:drug effects / Q000302:isolation & purification; D014364:Tryptophan / Q000737:chemistry
chemical_list: D000900:Anti-Bacterial Agents; D023181:Antimicrobial Cationic Peptides; D029941:Fish Proteins; D005452:Fluoresceins; C585732:NRC-16 peptide, Glyptocephalus cynoglossus; D014364:Tryptophan; C007740:fluorexon