514adda243a03e5c6b0bbb52a4afb04a intjoumolsci_a2018v19n8p2170.pdf 508b96686209fe362a4b311fbf79a816122fa3e8 intjoumolsci_a2018v19n8p2170.pdf 6128ec1db96ba353a01f85ed16bac4fe8cdab9713a5fda606ba734d2e9793ec0 intjoumolsci_a2018v19n8p2170.pdf Title: Unveiling the Multifaceted Mechanisms of Antibacterial Activity of Buforin II and Frenatin 2.3S Peptides from Skin Micro-Organs of the Orinoco Lime Treefrog (Sphaenorhynchus lacteus) Subject: Amphibian skin is a rich source of natural compounds with diverse antimicrobial and immune defense properties. Our previous studies showed that the frog skin secretions obtained by skin micro-organs from various species of Colombian anurans have antimicrobial activities against bacteria and viruses. We purified for the first time two antimicrobial peptides from the skin micro-organs of the Orinoco lime treefrog (Sphaenorhynchus lacteus) that correspond to Buforin II (BF2) and Frenatin 2.3S (F2.3S). Here, we have synthesized the two peptides and tested them against Gram-negative and Gram-positive bacteria, observing an effective bactericidal activity at micromolar concentrations. Evaluation of BF2 and F2.3S membrane destabilization activity on bacterial cell cultures and synthetic lipid bilayers reveals a distinct membrane interaction mechanism. BF2 agglutinates E. coli cells and synthetic vesicles, whereas F2.3S shows a high depolarization and membrane destabilization activities. Interestingly, we found that F2.3S is able to internalize within bacterial cells and can bind nucleic acids, as previously reported for BF2. Moreover, bacterial exposure to both peptides alters the expression profile of genes related to stress and resistance response. Overall, these results show the multifaceted mechanism of action of both antimicrobial peptides that can provide alternative tools in the fight against bacterial resistance. Keywords: antibacterial activity; antimicrobial peptides; bacterial agglutination; buforin II; DNA binding; frenatin 2.3S; frog skin secretions; membrane translocation; membrane leakage Author: Carolina Muñoz-Camargo, Vivian A. Salazar, Laura Barrero-Guevara, Sandra Camargo, Angela Mosquera, Helena Groot and Ester Boix Creator: LaTeX with hyperref package Producer: pdfTeX-1.40.18 CreationDate: Wed Jul 25 08:33:20 2018 CEST ModDate: Fri Oct 19 11:58:38 2018 CEST Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 21 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 2615756 bytes Optimized: yes PDF version: 1.5 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- KORLVE+VnURWPalladioL Type 1 Custom yes yes no 859 0 JDNTBR+URWPalladioL-Ital Type 1 Custom yes yes no 863 0 IEKFYN+URWPalladioL-Bold Type 1 Custom yes yes no 866 0 ZDBELT+URWPalladioL-BoldItal Type 1 Custom yes yes no 869 0 PVUNNK+URWPalladioL-Roma Type 1 Custom yes yes no 872 0 OXBKYU+TeXGyreAdventor-Bold Type 1 Custom yes yes no 876 0 IRWHZX+TimesNewRomanPSMT Type 1C WinAnsi yes yes no 880 0 CKUPDE+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 882 0 MCBCCI+SourceSansRoman-Regular Type 1C Custom yes yes no 891 0 MCBCCM+SourceSansRoman.613wght Type 1C Custom yes yes no 894 0 MCBCCO+SourceSansRoman-Bold Type 1C Custom yes yes no 897 0 REGJRM+EURM10 Type 1 Builtin yes yes no 543 0 ABCDEE+Palatino Linotype TrueType WinAnsi yes yes no 156 0 ABCDEE+Palatino Linotype,Italic TrueType WinAnsi yes yes no 159 0 ABCDEE+Palatino Linotype,Bold TrueType WinAnsi yes yes no 162 0 Times New Roman TrueType WinAnsi no no no 165 0 ABCDEE+Palatino Linotype CID TrueType Identity-H yes yes yes 171 0 ABCDEE+Palatino Linotype TrueType WinAnsi yes yes no 177 0 ABCDEE+Palatino Linotype,Italic TrueType WinAnsi yes yes no 180 0 ABCDEE+Palatino Linotype,Bold TrueType WinAnsi yes yes no 183 0 Times New Roman TrueType WinAnsi no no no 186 0 ABCDEE+Palatino Linotype CID TrueType Identity-H yes yes yes 192 0 QMDDXF+CMSY10 Type 1 Builtin yes yes no 547 0 ABCDEE+Palatino Linotype TrueType WinAnsi yes yes no 224 0 ABCDEE+Palatino Linotype,Italic TrueType WinAnsi yes yes no 227 0 ABCDEE+Palatino Linotype,Bold TrueType WinAnsi yes yes no 230 0 Times New Roman TrueType WinAnsi no no no 233 0 ABCDEE+Palatino Linotype CID TrueType Identity-H yes yes yes 239 0 Times New Roman,Bold TrueType WinAnsi no no no 242 0 ABCDEE+Palatino Linotype TrueType WinAnsi yes yes no 255 0 ABCDEE+Palatino Linotype,Italic TrueType WinAnsi yes yes no 258 0 ABCDEE+Palatino Linotype,Bold TrueType WinAnsi yes yes no 261 0 Times New Roman TrueType WinAnsi no no no 264 0 ABCDEE+Palatino Linotype TrueType WinAnsi yes yes no 270 0 ABCDEE+Palatino Linotype,Italic TrueType WinAnsi yes yes no 273 0 ABCDEE+Palatino Linotype,Bold TrueType WinAnsi yes yes no 276 0 Times New Roman TrueType WinAnsi no no no 279 0 ABCDEE+Palatino Linotype TrueType WinAnsi yes yes no 291 0 ABCDEE+Palatino Linotype,Italic TrueType WinAnsi yes yes no 294 0 ABCDEE+Palatino Linotype,Bold TrueType WinAnsi yes yes no 297 0 Times New Roman TrueType WinAnsi no no no 300 0 ABCDEE+Palatino Linotype CID TrueType Identity-H yes yes yes 306 0 JBTTSN+VnURWPalladioL-Italic Type 1 Custom yes yes no 551 0 MPUDFL+PazoMath Type 1 Builtin yes yes no 552 0 Jhove (Rel. 1.6, 2011-01-04) Date: 2018-10-26 04:46:18 CEST RepresentationInformation: intjoumolsci_a2018v19n8p2170.pdf ReportingModule: BYTESTREAM, Rel. 1.3 (2007-04-10) LastModified: 2018-10-19 11:58:58 CEST Size: 2615756 Format: bytestream Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/octet-stream Checksum: 70504d3b Type: CRC32 Checksum: 514adda243a03e5c6b0bbb52a4afb04a Type: MD5 Checksum: 508b96686209fe362a4b311fbf79a816122fa3e8 Type: SHA-1