Solving the alpha-conotoxin folding problem: Efficient selenium-directed on-resin generation of more potent and stable nicotinic acetylcholine receptor antaqonists

Muttenthaler, Marcus, Nevin, Simon T., Grishin, Anton A., Ngo, Shyuan T., Choy, Peng T., Daly, Norelle L., Hu, Shu-Hong, Armishaw, Christopher J., Wang, Ching-I. A., Lewis, Richard J., Martin, Jennifer L., Noakes, Peter G., Craik, David J., Adams, David J. and Alewood, Paul F. (2010) Solving the alpha-conotoxin folding problem: Efficient selenium-directed on-resin generation of more potent and stable nicotinic acetylcholine receptor antaqonists. Journal of the American Chemical Society, 132 10: 3514-3522. doi:10.1021/ja910602h


Author Muttenthaler, Marcus
Nevin, Simon T.
Grishin, Anton A.
Ngo, Shyuan T.
Choy, Peng T.
Daly, Norelle L.
Hu, Shu-Hong
Armishaw, Christopher J.
Wang, Ching-I. A.
Lewis, Richard J.
Martin, Jennifer L.
Noakes, Peter G.
Craik, David J.
Adams, David J.
Alewood, Paul F.
Title Solving the alpha-conotoxin folding problem: Efficient selenium-directed on-resin generation of more potent and stable nicotinic acetylcholine receptor antaqonists
Formatted title
Solving the α-conotoxin folding problem: Efficient selenium-directed on-resin generation of more potent and stable nicotinic acetylcholine receptor antaqonists

Journal name Journal of the American Chemical Society   Check publisher's open access policy
ISSN 0002-7863
1520-5126
1943-2984
Publication date 2010-03-17
Sub-type Article (original research)
DOI 10.1021/ja910602h
Volume 132
Issue 10
Start page 3514
End page 3522
Total pages 9
Place of publication Washington, DC, United States
Publisher American Chemical Society
Collection year 2011
Language eng
Subject C1
92 Health
11 Medical and Health Sciences
Formatted abstract
α-Conotoxins are tightly folded miniproteins that antagonize nicotinic acetylcholine receptors (nAChR) with high specificity for diverse subtypes. Here we report the use of selenocysteine in a supported phase method to direct native folding and produce α-conotoxins efficiently with improved biophysical properties. By replacing complementary cysteine pairs with selenocysteine pairs on an amphiphilic resin, we were able to chemically direct all five structural subclasses of α-conotoxins exclusively into their native folds. X-ray analysis at 1.4 Å resolution of α-selenoconotoxin PnIA confirmed the isosteric character of the diselenide bond and the integrity of the α-conotoxin fold. The α-selenoconotoxins exhibited similar or improved potency at rat diaphragm muscle and α3β4, α7, and α1β1δγ nAChRs expressed in Xenopus oocytes plus improved disulfide bond scrambling stability in plasma. Together, these results underpin the development of more stable and potent nicotinic antagonists suitable for new drug therapies, and highlight the application of selenocysteine technology more broadly to disulfide-bonded peptides and proteins.
Copyright © 2011 American Chemical Society

Keyword Native chemical ligation
Crystal structure
Glutathione peroxidase
Refined structure
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

 
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Created: Sun, 04 Apr 2010, 00:02:50 EST