Rapid Access to ω-Conotoxin Chimeras using Native Chemical Ligation
Gene Hopping A , Richard J. Lewis A and Paul F. Alewood A BA Institute for Molecular Bioscience, The University of Queensland, St Lucia, Qld 4072, Australia.
B Corresponding author. Email: P.Alewood@imb.uq.edu.au
Australian Journal of Chemistry 62(10) 1333-1338 https://doi.org/10.1071/CH09216
Submitted: 14 April 2009 Accepted: 9 June 2009 Published: 13 October 2009
Abstract
Grafting different regions of related peptides together to form a single protein chimera is a valuable tool in rapidly elucidating regions of activity or selectivity in peptides and proteins. To conveniently evaluate the contributions of the N- and C-terminal segments of ω-conotoxins CVID and MVIIC to activity, we employed native chemical ligation in CVID-MVIIC chimera design. Assembly of these peptide segments via the ligation method improved overall yield and coupling efficiency, with no difficult sequences encountered in contrast to the traditional full-length chain assembly of CVID. Radio-ligand binding assays revealed regions of importance for receptor recognition.
Acknowledgements
This work was supported by grants from the National Health & Medical Research Council Program Grant and the Australian Research Council.
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