Chemical Synthesis of A Pore‐Forming Antimicrobial Protein,Caenopore‐5, by Using Native Chemical Ligation at a Glu‐Cys Site |
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| Authors: | Karima Medini Dr Paul W R Harris Kiel Hards Dr Andrew J Dingley Gregory M Cook Prof Margaret A Brimble |
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| Affiliation: | 1. Maurice Wilkins Centre for Molecular Biodiscovery, School of Biological Sciences, The University of Auckland, 3A Symonds Street, Auckland 1010 (New Zealand);2. School of Chemical Sciences, The University of Auckland, 23 Symonds Street, Auckland 1010 (New Zealand);3. Department of Microbiology and Immunology, School of Medical Sciences, University of Otago, 720 Cumberland Street, Dunedin 9054 (New Zealand);4. Institute of Complex Systems 6, Structural Biochemistry, Forschungszentrum Jülich, 52425 Jülich (Germany) |
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| 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 1H 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. |
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| Keywords: | antibiotics antimicrobial protein caenopore native chemical ligation protein design protein folding |
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