C-terminal amidation of PMAP-23: translocation to the inner membrane of Gram-negative bacteria.
PMAP-23 is a member of the cathelicidin family derived from pig myeloid cells and has potent antimicrobial activity. Amidation of the carboxyl terminus (C-terminus) of an antimicrobial peptide generally enhances its structural stability and antimicrobial activity or decreases its cytotoxicity. The aim of the present study was to investigate the effect of amidation on the mode of action in PMAP-23. Irrespective of amidation, PMAP-23 adopts a helix-hinge-helix structure in a membrane-mimetic environment. The antibacterial activities of PMAP-23C, which had a free C-terminus, and PMAP-23N, which had an amidated C-terminus, were similar against Gram-negative bacteria, reflecting a similar ability to neutralize lipopolysaccharide. However, PMAP-23N assumed a perpendicular orientation across the outer to the inner leaflet of the bacterial inner membrane, while PMAP-23C was orientated parallel to the lipid bilayer, as determined by following the blue shift in tryptophan fluorescence, as well as calcein release from liposomes and SYTOX Green uptake assays. These results suggest that N-terminal amidation of PMAP-23 provides structural stability and increases the peptide's cationic charge, facilitating translocation into the bacterial inner membrane.
Research Center for Proteineous Materials (RCPM), Chosun University, Kwangju, Korea.
Jin-Young
JY
Kim
Seong-Cheol
SC
Park
Moon-Young
MY
Yoon
Kyung-Soo
KS
Hahm
Yoonkyung
Y
Park
D000900:Anti-Bacterial Agents / Q000737:chemistry* / Q000378:metabolism* / Q000494:pharmacology; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry* / Q000378:metabolism* / Q000494:pharmacology; D001692:Biological Transport; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism*; D006090:Gram-Negative Bacteria / Q000187:drug effects / Q000378:metabolism*; D006801:Humans
D000900:Anti-Bacterial Agents; D023181:Antimicrobial Cationic Peptides; C086301:porcine myeloid antibacterial peptide 23