Efficient enzymatic cyclization of an inhibitory cystine knot-containing peptide

Kwon, Soohyun, Bosmans, Frank, Kaas, Quentin, Cheneval, Olivier, Conibear, Anne C., Rosengren, K. Johan, Wang, Conan K., Schroeder, Christina I. and Craik, David J. (2016) Efficient enzymatic cyclization of an inhibitory cystine knot-containing peptide. Biotechnology and Bioengineering, 113 10: 2202-2212. doi:10.1002/bit.25993


Author Kwon, Soohyun
Bosmans, Frank
Kaas, Quentin
Cheneval, Olivier
Conibear, Anne C.
Rosengren, K. Johan
Wang, Conan K.
Schroeder, Christina I.
Craik, David J.
Title Efficient enzymatic cyclization of an inhibitory cystine knot-containing peptide
Journal name Biotechnology and Bioengineering   Check publisher's open access policy
ISSN 1097-0290
0006-3592
Publication date 2016-10-01
Year available 2016
Sub-type Article (original research)
DOI 10.1002/bit.25993
Open Access Status Not yet assessed
Volume 113
Issue 10
Start page 2202
End page 2212
Total pages 11
Place of publication Hoboken, United States
Publisher John Wiley and Sons
Language eng
Formatted abstract
Disulfide-rich peptides isolated from cone snails are of great interest as drug leads due to their high specificity and potency toward therapeutically relevant ion channels and receptors. They commonly contain the inhibitor cystine knot (ICK) motif comprising three disulfide bonds forming a knotted core. Here we report the successful enzymatic backbone cyclization of an ICK-containing peptide κ-PVIIA, a 27-amino acid conopeptide from Conus purpurascens, using a mutated version of the bacterial transpeptidase, sortase A. Although a slight loss of activity was observed compared to native κ-PVIIA, cyclic κ-PVIIA is a functional peptide that inhibits the Shaker voltage-gated potassium (Kv) channel. Molecular modeling suggests that the decrease in potency may be related to the loss of crucial, but previously unidentified electrostatic interactions between the N-terminus of the peptide and the Shaker channel. This hypothesis was confirmed by testing an N-terminally acetylated κ-PVIIA, which shows a similar decrease in activity. We also investigated the conformational dynamics and hydrogen bond network of cyc-PVIIA, both of which are important factors to be considered for successful cyclization of peptides. We found that cyc-PVIIA has the same conformational dynamics, but different hydrogen bond network compared to those of κ-PVIIA. The ability to efficiently cyclize ICK peptides using sortase A will enable future protein engineering for this class of peptides and may help in the development of novel therapeutic molecules. Biotechnol. Bioeng. 2016;113: 2202–2212.
Keyword Cyclization
Inhibitory cystine knot
Sortase A
κ-PVIIA
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: HERDC Pre-Audit
School of Biomedical Sciences Publications
Institute for Molecular Bioscience - Publications
 
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Created: Wed, 07 Sep 2016, 22:57:29 EST by Susan Allen on behalf of Learning and Research Services (UQ Library)