title: Chemical synthesis of a pore-forming antimicrobial protein, caenopore-5, by using native chemical ligation at a glu-cys site.
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.
authors.affiliation: Maurice Wilkins Centre for Molecular Biodiscovery, School of Biological Sciences, The University of Auckland, 3A Symonds Street, Auckland 1010 (New Zealand).
authors.forename: Karima
authors.initials: K
authors.lastname: Medini
authors.forename: Paul W R
authors.initials: PW
authors.lastname: Harris
authors.forename: Kiel
authors.initials: K
authors.lastname: Hards
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Dingley
authors.forename: Gregory M
authors.initials: GM
authors.lastname: Cook
authors.forename: Margaret A
authors.initials: MA
authors.lastname: Brimble
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000890:Anti-Infective Agents / Q000138:chemical synthesis* / Q000737:chemistry; D017173:Caenorhabditis elegans / Q000378:metabolism*; D029742:Caenorhabditis elegans Proteins / Q000138:chemical synthesis / Q000737:chemistry* / Q000235:genetics / Q000494:pharmacology; D002462:Cell Membrane / Q000187:drug effects; D060326:Chemistry Techniques, Synthetic; D002942:Circular Dichroism; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D010539:Permeability; D017510:Protein Folding; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D060327:Solid-Phase Synthesis Techniques
chemical_list: D000890:Anti-Infective Agents; D029742:Caenorhabditis elegans Proteins; D011994:Recombinant Proteins
keywords: antibiotics; antimicrobial protein; caenopore; native chemical ligation; protein design; protein folding