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Pleurocidin, an α-helical cationic antimicrobialpeptide, was isolated from skin mucosa of winter flounder(Pleuronectes americanus). It had strong antimicrobial activitiesagainst Gram-positive and Gram-negative bacteria, but hadvery weak hemolytic activity. The Gly13,17→Ala analoghad dramatically increased hemolytic activity. The bacterialcel selectivity of pleurocidin was confirmed through themembrane-disrupting and membrane-binding affinities usingdye leakage, tryptophan fluorescence blue shift, and tryptophanquenching experiments. However, the non-cel-selectiveantimicrobial peptide, pleurocidin-AA, interacts strongly withboth negatively charged and zwitterionic phospholipid membranes,the latter of which are the major constituents of the outerleaflet of erythrocytes. Circular dihroism spectra showedthat pleurocidin-AA has much higher contents of α-helicalconformation than pleurocidin. The tertiary structure determinedby NMR spectroscopy showed that pleurocidin has a flexiblestructure between the long helix from Gly3 to Gly17 and theshort helix from Gly17 to Leu25. Cel-selective antimicrobialpeptide pleurocidin interacts strongly with negatively chargedphospholipid membranes, which mimic bacterial membranes.Structural flexibility between the two helices may play a keyrole in bacterial cel selectivity of pleurocidin.

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