Tuning of a Membrane-Perforating Antimicrobial Peptide to Selectively Target Membranes of Different Lipid Composition

Charles H. Chen*, Charles G. Starr, Shantanu Guha, William C. Wimley, Martin B. Ulmschneider, Jakob P. Ulmschneider

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

13 Citations (Scopus)

Abstract

The use of designed antimicrobial peptides as drugs has been impeded by the absence of simple sequence-structure–function relationships and design rules. The likely cause is that many of these peptides permeabilize membranes via highly disordered, heterogeneous mechanisms, forming aggregates without well-defined tertiary or secondary structure. We suggest that the combination of high-throughput library screening with atomistic computer simulations can successfully address this challenge by tuning a previously developed general pore-forming peptide into a selective pore-former for different lipid types. A library of 2916 peptides was designed based on the LDKA template. The library peptides were synthesized and screened using a high-throughput orthogonal vesicle leakage assay. Dyes of different sizes were entrapped inside vesicles with varying lipid composition to simultaneously screen for both pore size and affinity for negatively charged and neutral lipid membranes. From this screen, nine different LDKA variants that have unique activity were selected, sequenced, synthesized, and characterized. Despite the minor sequence changes, each of these peptides has unique functional properties, forming either small or large pores and being selective for either neutral or anionic lipid bilayers. Long-scale, unbiased atomistic molecular dynamics (MD) simulations directly reveal that rather than rigid, well-defined pores, these peptides can form a large repertoire of functional dynamic and heterogeneous aggregates, strongly affected by single mutations. Predicting the propensity to aggregate and assemble in a given environment from sequence alone holds the key to functional prediction of membrane permeabilization. Graphic Abstract: [Figure not available: see fulltext.]

Original languageEnglish
Pages (from-to)75-96
Number of pages22
JournalJournal of Membrane Biology
Volume254
Issue number1
DOIs
Publication statusPublished - Feb 2021

Keywords

  • Antimicrobial peptides
  • Bacterial selectivity
  • Drug-resistant bacteria
  • Leucine-rich peptide
  • Pore formation
  • Protein folding

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