title: Antimicrobial characterization of site-directed mutagenesis of porcine beta defensin 2.
abstract: Porcine β defensin 2 (pBD2) is a small, cationic and amphiphilic antimicrobial peptide. It has broad antimicrobial activities against bacteria and plays an important role in host defense. In order to enhance its antimicrobial activity and better understand the effect of positively charged residues on its activity, we substituted eight amino acid residues with arginine or lysine respectively. All mutants were cloned and expressed in BL21 (DE3) plysS and the mutant proteins were then purified. These mutant versions had higher positive charges but similar structural configurations compared to the wild-type pBD2. Moreover, these mutant proteins showed different antimicrobial activities against E. coli and S. aureus. The mutant I4R of pBD2 had the highest antimicrobial activity. In addition, all the mutants showed low hemolytic activities. Our results indicated that the positively charged residues were not the only factor that influenced antimicrobial activity, but other factors such as distribution of these residues on the surface of defensins might also contribute to their antimicrobial potency.
authors.affiliation: Department of Animal and Veterinary Science, Henan Agricultural University, Zhengzhou, Henan Province, The People's Republic of China.
authors.forename: Xian-xian
authors.initials: XX
authors.lastname: Huang
authors.affiliation: Department of Animal and Veterinary Science, Henan Agricultural University, Zhengzhou, Henan Province, The People's Republic of China.
authors.forename: Chun-yu
authors.initials: CY
authors.lastname: Gao
authors.affiliation: Department of Animal and Veterinary Science, Henan Agricultural University, Zhengzhou, Henan Province, The People's Republic of China.
authors.forename: Qing-jun
authors.initials: QJ
authors.lastname: Zhao
authors.affiliation: Department of Animal and Veterinary Science, Henan Agricultural University, Zhengzhou, Henan Province, The People's Republic of China.
authors.forename: Chun-li
authors.initials: CL
authors.lastname: Li
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000494:pharmacology* / Q000633:toxicity; D004926:Escherichia coli / Q000187:drug effects; D006461:Hemolysis; D008969:Molecular Sequence Data; D017354:Point Mutation; D013211:Staphylococcus aureus / Q000187:drug effects; D013552:Swine; D023083:beta-Defensins / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology* / Q000633:toxicity
chemical_list: D000900:Anti-Bacterial Agents; D023083:beta-Defensins