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Quantifying the Antifungal Activity of Peptides Against Candida albicans
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Evolution-Based Protein Engineering for Antifungal Peptide Improvement.

Jing Gu1,2, Noriyoshi Isozumi3, Shouli Yuan1,2

  • 1Group of Peptide Biology and Evolution, State Key Laboratory of Integrated Management of Pest Insects and Rodents, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.

Molecular Biology and Evolution
|July 28, 2021
PubMed
Summary
This summary is machine-generated.

Researchers engineered antimicrobial peptides (AMPs) to combat fungal infections. An evolution-based strategy identified a key site (UEAMS) that significantly boosted antifungal activity, offering a promising alternative to conventional antibiotics.

Keywords:
Candida albicansCremycin-5antimicrobial peptideepistasisparaloguniversally enhanceable activity-modulating site (UEAMS)

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Drug Discovery

Background:

  • Antimicrobial peptides (AMPs) are promising antibiotic alternatives due to low resistance.
  • Clinical application of AMPs is limited by low activity, high cost, and cytotoxicity.
  • Engineering AMPs is challenging due to complex sequence-structure-function relationships.

Purpose of the Study:

  • To develop an evolution-based strategy for enhancing antifungal activity of AMPs.
  • To identify and characterize sites responsible for improved antifungal efficacy.
  • To engineer a nematode-sourced defensin (Cremycin-5) for increased potency and stability.

Main Methods:

  • Sequence-activity comparison between Cremycin-5 and paralogs to identify key sites.
  • Saturation mutagenesis at identified sites to screen for enhanced activity.
  • Antifungal activity assays against multiple fungal species, including Candida albicans clinical isolates.
  • Molecular dynamic simulations to elucidate the mechanism of action.
  • Analysis of sequence, structure, and epistasis for evolutionary insights.

Main Results:

  • Identified a Universally Enhanceable Activity-Modulating Site (UEAMS) at position Glu-15.
  • Mutations at Glu-15 universally enhanced antifungal activity against multiple fungal species.
  • The Glu15Lys mutant showed >9-fold increased potency against Candida albicans, inhibiting cytokinesis.
  • The engineered mutant exhibited high stability, rapid killing kinetics, and no detectable hemolysis.
  • Molecular dynamics suggested allosteric regulation of distant functional residues via UEAMS mutations.

Conclusions:

  • The evolution-based strategy successfully engineered a potent antifungal AMP.
  • UEAMS represents a novel target for improving AMP efficacy and overcoming limitations.
  • The findings provide a new pathway for developing AMP-based antifungal drugs.
  • Understanding epistatic interactions can guide future AMP engineering efforts.