title: Antimicrobial activity of omwaprin, a new member of the waprin family of snake venom proteins.
abstract: We have isolated and characterized omwaprin, a 50-amino-acid cationic protein from the venom of inland taipan (Oxyuranus microlepidotus). It is a new member of the waprin family of snake venom proteins. A synthetic gene was designed and constructed for expressing the recombinant protein in Escherichia coli. Recombinant omwaprin was used for carrying out functional analyses. The protein is non-toxic to Swiss albino mice at doses of up to 10 mg/kg when administered intraperitoneally. However, it shows selective and dose-dependant antibacterial activity against Gram-positive bacteria. The minimum inhibitory doses were in the range 2-10 microg for selected species of bacteria in radial diffusion assays. The antibacterial activity is salt-tolerant up to 350 mM NaCl. However, omwaprin lost its antibacterial activity upon reduction and alkylation of its cysteine residues, or upon deletion of six N-terminal amino acid residues, four of which are positively charged. These observations indicate that the three-dimensional structure constrained by four disulfide bonds and the N-terminal residues are essential for its activity. The mechanism of action is via membrane disruption, as shown by scanning electron microscopy. Importantly, omwaprin lacks haemolytic activity on human erythrocytes. This demonstrates the specificity of omwaprin for bacterial membranes. Unlike other reported WAP (whey acidic protein) domain-containing antibacterial proteins, including elafin, EPPIN (epididymal proteinase inhibitor), SWAM1 and SWAM2 [single WAP (whey acidic protein) motif proteins 1 and 2] and SLPI (secretory leucocyte proteinase inhibitor), omwaprin shows species-specific activity on the Gram-positive bacteria tested.
authors.affiliation: Department of Biological Sciences, Faculty of Science, National University of Singapore, Singapore 117543.
authors.forename: Dileep G
authors.initials: DG
authors.lastname: Nair
authors.forename: Bryan G
authors.initials: BG
authors.lastname: Fry
authors.forename: Paul
authors.initials: P
authors.lastname: Alewood
authors.forename: Prakash P
authors.initials: PP
authors.lastname: Kumar
authors.forename: R Manjunatha
authors.initials: RM
authors.lastname: Kini
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000890:Anti-Infective Agents / Q000737:chemistry* / Q000494:pharmacology*; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry* / Q000235:genetics / Q000494:pharmacology*; D001483:Base Sequence; D003001:Cloning, Molecular; D004546:Elapid Venoms / Q000737:chemistry* / Q000235:genetics / Q000494:pharmacology / Q000633:toxicity; D017815:Elapidae / Q000378:metabolism; D006801:Humans; D008297:Male; D051379:Mice; D008855:Microscopy, Electron, Scanning; D008969:Molecular Sequence Data; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D021241:Spectrometry, Mass, Electrospray Ionization; D013329:Structure-Activity Relationship
chemical_list: D000890:Anti-Infective Agents; D023181:Antimicrobial Cationic Peptides; D004546:Elapid Venoms; D011506:Proteins; D011994:Recombinant Proteins; C517856:omwaprin protein, Oxyuranus microlepidotus