Efficient backbone cyclization of linear peptides by a recombinant asparaginyl endopeptidase

Harris, Karen S., Durek, Thomas, Kaas, Quentin, Poth, Aaron G., Gilding, Edward K., Conlan, Brendon F., Saska, Ivana, Daly, Norelle, Van Der Weerden, Nicole L., Craik, David J., and Anderson, Marilyn A, (2015) Efficient backbone cyclization of linear peptides by a recombinant asparaginyl endopeptidase. Nature Communications, 6. 10199.

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Cyclotides are diverse plant backbone cyclized peptides that have attracted interest as pharmaceutical scaffolds, but fundamentals of their biosynthetic origin remain elusive. Backbone cyclization is a key enzyme-mediated step of cyclotide biosynthesis and confers a measure of stability on the resultant cyclotide. Furthermore, cyclization would be desirable for engineered peptides. Here we report the identification of four asparaginyl endopeptidases (AEPs), proteases implicated in cyclization, from the cyclotide-producing plant Oldenlandia affinis. We recombinantly express OaAEP1 b and find it functions preferably as a cyclase by coupling C-terminal cleavage of propeptide substrates with backbone cyclization. Interestingly, OaAEP1 b cannot cleave at the N-terminal site of O. affinis cyclotide precursors, implicating additional proteases in cyclotide biosynthesis. Finally, we demonstrate the broad utility of this enzyme by cyclization of peptides unrelated to cyclotides. We propose that recombinant OaAEP1 b is a powerful tool for use in peptide engineering applications where increased stability of peptide products is desired.

Item ID: 44446
Item Type: Article (Research - C1)
ISSN: 2041-1723
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Funders: Australian Research Council (ARC)
Projects and Grants: ARC DP0984390 , ARC DP150100443
Date Deposited: 17 Jun 2016 03:48
FoR Codes: 11 MEDICAL AND HEALTH SCIENCES > 1101 Medical Biochemistry and Metabolomics > 110106 Medical Biochemistry: Proteins and Peptides (incl Medical Proteomics) @ 100%
SEO Codes: 97 EXPANDING KNOWLEDGE > 970111 Expanding Knowledge in the Medical and Health Sciences @ 100%
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