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Chemical Synthesis of A Pore‐Forming Antimicrobial Protein, Caenopore‐5, by Using Native Chemical Ligation at a Glu‐Cys Site
Author(s) -
Medini Karima,
Harris Paul W. R.,
Hards Kiel,
Dingley Andrew J.,
Cook Gregory M.,
Brimble Margaret A.
Publication year - 2015
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201402513
Subject(s) - native chemical ligation , antimicrobial , circular dichroism , thioester , chemical ligation , chemistry , peptide , solid phase synthesis , antimicrobial peptides , biochemistry , combinatorial chemistry , recombinant dna , ligation , chemical synthesis , biology , organic chemistry , microbiology and biotechnology , in vitro , enzyme , gene
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.