Chemical Synthesis of a Pore-Forming Antimicrobial Protein, caenopore-5, by Using Native Chemical Ligation at a Glu-Cys Site

Chembiochem. 2015 Jan 19;16(2):328-36. doi: 10.1002/cbic.201402513. Epub 2014 Nov 25.

Abstract

The 2014 report from the World Health Organization (WHO) on antimicrobial resistance revealed an alarming rise in antibiotic resistance all around the world. Unlike classical antibiotics, with the exception of a few species, no acquired resistance towards antimicrobial peptides (AMPs) has been reported. Therefore, AMPs represent leads for the development of novel antibiotics. Caenopore-5 is constitutively expressed in the intestine of the nematode Caenorhabditis elegans and is a pore-forming AMP. The protein (82 amino acids) was successfully synthesised by using Boc solid-phase peptide synthesis and native chemical ligation. No γ-linked by-product was observed despite the use of a C-terminal Glu-thioester. The folding of the synthetic protein was confirmed by (1) H NMR spectroscopy and circular dichroism and compared with data recorded for recombinant caenopore-5. The permeabilisation activities of the protein and of shortened analogues were evaluated.

Keywords: antibiotics; antimicrobial protein; caenopore; native chemical ligation; protein design; protein folding.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Anti-Infective Agents / chemical synthesis*
  • Anti-Infective Agents / chemistry
  • Caenorhabditis elegans / metabolism*
  • Caenorhabditis elegans Proteins / chemical synthesis
  • Caenorhabditis elegans Proteins / chemistry*
  • Caenorhabditis elegans Proteins / genetics
  • Caenorhabditis elegans Proteins / pharmacology
  • Cell Membrane / drug effects
  • Chemistry Techniques, Synthetic
  • Circular Dichroism
  • Magnetic Resonance Spectroscopy
  • Molecular Sequence Data
  • Permeability
  • Protein Folding
  • Protein Structure, Secondary
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Solid-Phase Synthesis Techniques

Substances

  • Anti-Infective Agents
  • Caenorhabditis elegans Proteins
  • Recombinant Proteins