Identification of novel Amurin-2 variants from the skin secretion of Rana amurensis, and the design of cationicity-enhanced analogues

Biochem Biophys Res Commun. 2018 Mar 18;497(4):943-949. doi: 10.1016/j.bbrc.2018.01.124. Epub 2018 Jan 31.

Abstract

Rana amurensis is important in Chinese medicine as its skin secretions contain abundant bioactive peptides. Here, we have identified the antimicrobial peptide Amurin-2 and three highly-conserved variants, Amurin-2a, Amurin-2b and Amurin-2c through a combination of molecular cloning and MS/MS fragmentation sequencing. Synthetic replicates of these peptides demonstrate potent antimicrobial activity against S. aureus, whilst some have activity against C.albicans and even resistant bacterial MRSA. Furthermore, two Lys-analogues (K4-Amurin-2 and K11-Amurin-2) were designed to improve the bioactive function and the antimicrobial activity of K4-Amurin-2 against E.coli was enhanced distinctly. In addition, the two modified peptides also showed more potent activity against S. aureus, C. albicans and MRSA strains. Meanwhile, these peptides showed inhibitory effect on the cell viability of several cancer cells. As a result, these structural and functional studies of Amurin-2 variants and analogues could provide insights for future antimicrobial peptide design.

Keywords: Amphibian skin secretion; Antimicrobial peptide; Cationicity-related peptide design.

MeSH terms

  • Amphibian Proteins / genetics*
  • Animals
  • Anti-Bacterial Agents / pharmacology
  • Antimicrobial Cationic Peptides / chemical synthesis*
  • Antimicrobial Cationic Peptides / pharmacology
  • Candida albicans / drug effects
  • Cell Line, Tumor
  • Cell Survival / drug effects
  • Cloning, Molecular
  • Drug Design
  • Genetic Variation
  • Humans
  • Methicillin-Resistant Staphylococcus aureus / drug effects
  • Neoplasms / drug therapy
  • Neoplasms / pathology
  • Ranidae / genetics*
  • Skin / metabolism*
  • Staphylococcus aureus / drug effects
  • Tandem Mass Spectrometry

Substances

  • Amphibian Proteins
  • Anti-Bacterial Agents
  • Antimicrobial Cationic Peptides