The role of the central L- or D-Pro residue on structure and mode of action of a cell-selective alpha-helical IsCT-derived antimicrobial peptide.
IsCT-P (ILKKIWKPIKKLF-NH2) is a novel alpha-helical antimicrobial peptide with bacterial cell selectivity designed from a scorpion-derived peptide IsCT. To investigate the role of L- or D-Pro kink on the structure and the mode of action of a short alpha-helical antimicrobial peptide with bacterial cell selectivity, we synthesized IsCT-p, in which D-Pro is substituted for L-Pro8 of IsCT-P. CD spectra revealed that IsCT-P adopted a typical alpha-helical structure in various membrane-mimicking conditions, whereas IsCT-p showed a random structure. This result indicated that D-Pro in the central position of a short alpha-helical peptide provides more remarkable structural flexibility than L-Pro. Despite its higher antibacterial activity, IsCT-p was much less effective at inducing dye leakage in the negatively charged liposome mimicking bacterial membrane and induced no or little membrane potential depolarization of Staphylococcus aureus. Confocal laser scanning microscopy showed that IsCT-p penetrated the bacterial cell membrane and accumulated in the cytoplasm, whereas IsCT-P remained outside or on the cell membrane. These results suggested that the major target of IsCT-P and IsCT-p is the bacterial membranes and intracellular components, respectively. Collectively, our results demonstrated that the central D-Pro kink in alpha-helical antimicrobial peptides plays an important role in penetrating bacterial membrane as well as bacterial cell selectivity.
Lim
Shin Saeng
SS
Department of Bio-Materials, Graduate School and Research Center for Proteineous Materials, Chosun University, Gwangju 501-759, Republic of Korea.
Kim
Yangmee
Y
Park
Yoonkyung
Y
Kim
Jae Il
JI
Park
Il-Seon
IS
Hahm
Kyung-Soo
KS
Shin
Song Yub
SY
D023181:Antimicrobial Cationic Peptides / Q000737:chemistry* / Q000494:pharmacology*; D002462:Cell Membrane / Q000737:chemistry / Q000187:drug effects*; D002463:Cell Membrane Permeability / Q000187:drug effects*; D002478:Cells, Cultured; D004912:Erythrocytes / Q000187:drug effects; D004926:Escherichia coli / Q000166:cytology* / Q000187:drug effects*; D006461:Hemolysis / Q000187:drug effects; D006801:Humans; D007536:Isomerism; D008560:Membrane Fluidity / Q000187:drug effects; D012604:Scorpion Venoms / Q000737:chemistry* / Q000494:pharmacology*; D013329:Structure-Activity Relationship
D023181:Antimicrobial Cationic Peptides; C435485:IsCT protein, Opisthacanthus madagascariensis; C516429:IsCT-P peptide; D012604:Scorpion Venoms