Atypical alpha-conotoxin LtIA from Conus litteratus targets a novel microsite of the α3β2 nicotinic receptor

Luo, Sulan, Akondi, Kalyana Bharati, Zhangsun, Dongting, Wu, Yong, Zhu, Xiaopeng, Hu, Yuanyan, Christensen, Sean, Dowell, Cheryl, Daly, Norelle, Craik, David, Wang, Ching-I., Lewis, Richard J., Alewood, Paul F., and McIntosh, J. Michael (2010) Atypical alpha-conotoxin LtIA from Conus litteratus targets a novel microsite of the α3β2 nicotinic receptor. Journal of Biological Chemistry, 285 (16). pp. 12355-12366.

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Abstract

Different nicotinic acetylcholine receptor (nAChR) subtypes are implicated in learning, pain sensation, and disease states, including Parkinson disease and nicotine addiction. α-Conotoxins are among the most selective nAChR ligands. Mechanistic insights into the structure, function, and receptor interaction of α-conotoxins may serve as a platform for development of new therapies. Previously characterized α-conotoxins have a highly conserved Ser-Xaa-Pro motif that is crucial for potent nAChR interaction. This study characterized the novel α-conotoxin LtIA, which lacks this highly conserved motif but potently blocked α3β2 nAChRs with a 9.8 nm IC(50) value. The off-rate of LtIA was rapid relative to Ser-Xaa-Pro-containing alpha-conotoxin MII. Nevertheless, pre-block of α3β2 nAChRs with LtIA prevented the slowly reversible block associated with MII, suggesting overlap in their binding sites. nAChR beta subunit ligand-binding interface mutations were used to examine the >1000-fold selectivity difference of LtIA for α3β2 versus α3β4 nAChRs. Unlike MII, LtIA had a >900-fold increased IC(50) value on α3β2(F119Q) versus wild type nAChRs, whereas T59K and V111I β2 mutants had little effect. Molecular docking simulations suggested that LtIA had a surprisingly shallow binding site on the α3β2 nAChR that includes β2 Lys-79. The K79A mutant disrupted LtIA binding but was without effect on an LtIA analog where the Ser-Xaa-Pro motif is present, consistent with distinct binding modes.

Item ID: 27052
Item Type: Article (Research - C1)
ISSN: 1083-351X
Keywords: peptides; peptides/Chemical Synthesis; peptides/interactions; peptides/Neuropeptide; receptors; oxins; toxin/Channels
Date Deposited: 07 Jun 2013 04:12
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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