Origin of low mammalian cell toxicity in a class of highly active antimicrobialamphipathic helical peptides

A Hawrani, R.A Howe, T.R Walsh, C.E Dempsey

Research output: Contribution to journalArticle (Academic Journal)peer-review

67 Citations (Scopus)

Abstract

We recently described a novel antimicrobial peptide, RTA3, derived from the commensal organism Streptococcus mitis, with strong anti-Gram-negative activity, low salt-sensitivity, and minimal mammalian cell toxicity in vitro and in vivo. This peptide conforms to the positively-charged, amphipathic helical peptide motif, but has a positively charged amino acid (Arg5) on the non-polar face that forms when the peptide structures upon membrane binding. We surmised that disruption of the hydrophobic face with a positively-charged residue plays a role in minimizing eukaryotic cell toxicity, and tested this using a mutant with an Arg5->Leu5 substitution. The greatly enhanced toxicity in the mutant peptide correlated with its ability to bind and adopt helical conformations upon interacting with neutral membranes; the wild type peptide, RTA3 did not bind to neutral membranes (binding constant reduced by at least 1000-fold). Spectroscopic analysis indicates that disruption of the hydrophobic face of the parent peptide is accommodated in negatively charged membranes without partial peptide unfolding. These observations apply generally to amphipathic helical peptides of this class since we obtained similar results with a peptide and mutant pair [Chen et al. (2005) J. Biol. Chem 280, 12316-12329] having similar structural properties. In contrast to previous interpretations, we demonstrate that these peptides simply do not bind well to membranes (like those of eukaryotes) with exclusively neutral lipids in their external bilayer leaflet. We highlight a significant role for tryptophan in promoting binding of amphipathic helical peptides to neutral bilayers, augmenting the arsenal of strategies to reduce mammalian toxicity in antimicrobial peptides.
Translated title of the contributionOrigin of low mammalian cell toxicity in a class of highly active antimicrobialamphipathic helical peptides
Original languageEnglish
Pages (from-to)18636 - 18645
Number of pages10
JournalJournal of Biological Chemistry
Volume283
Issue number27
DOIs
Publication statusPublished - Jul 2008

Bibliographical note

Publisher: American Society for Biochemistry and Molecular Biology

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