Antimicrobial specificity and mechanism of action of disulfide-removed linear analogs of the plant-derived Cys-rich antimicrobial peptide Ib-AMP1.
Ib-AMP1 is a 20-residue disulfide-linked beta-sheet antimicrobial peptide found in the seeds of Impatiens balsamina. In order to investigate the effects of the 2 disulfide bonds on the antimicrobial specificity, to determine the mechanism of antimicrobial action of Ib-AMP1 and to develop novel cell-selective antimicrobial peptides with improved antimicrobial specificity as compared to wild-type Ib-AMP1, we synthesized a disulfide-removed linear analog of Ib-AMP1 with L-Pro, D-Pro or peptoid residues (Nala and Nlys) at the central position of the molecule. All linear analogs displayed a 3.7-4.8-fold higher antimicrobial specificity than wild-type Ib-AMP1, indicating that the disulfide bonds of Ib-AMP1 analogs are not essential for its antimicrobial specificity. Circular dichroism spectra revealed that the peptoid residues, as well as the proline at the central position of disulfide bond-removed Ib-AMP1 analogs, induce a beta-turn structure in a negatively charged bacterial membrane-mimicking environment. Ib-AMP1 was not effective in depolarizing the cytoplasmic membranes of Staphylococcus aureus and showed almost no leakage of calcein from negatively charged bacterial membranes mimicking lipid vesicles. In contrast, all linear analogs caused very weak dye leakage from negatively charged vesicles, but they almost completely depolarized the membrane potential of S. aureus cells. Collectively, our results suggest that the target of Ib-AMP1 may not be the cytoplasmic membranes of bacteria but their intracellular components. All linear analogs exhibit lethality due to their ability to form small channels that permit the transit of ions or protons and not molecules as large as calcein, and not by disrupting membranes.
Department of Bio-Materials, Graduate School, School of Medicine, Chosun University, Gwangju 501-759, Republic of Korea.
Peng
P
Wang
Jeong-Kyu
JK
Bang
Hak Jun
HJ
Kim
Jin-Kyoung
JK
Kim
Yangmee
Y
Kim
Song Yub
SY
Shin
D000595:Amino Acid Sequence; D000890:Anti-Infective Agents / Q000737:chemistry* / Q000494:pharmacology*; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry* / Q000494:pharmacology*; D002942:Circular Dichroism; D003545:Cysteine / Q000737:chemistry*; D004220:Disulfides / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D008564:Membrane Potentials / Q000187:drug effects; D008969:Molecular Sequence Data; D013211:Staphylococcus aureus / Q000187:drug effects* / Q000378:metabolism
D000890:Anti-Infective Agents; D023181:Antimicrobial Cationic Peptides; D004220:Disulfides; D003545:Cysteine