Isolation and characterization of α-conotoxin LsIA with potent activity at nicotinic acetylcholine receptors

Marco C Inserra; Shiva N Kompella; Irina Vetter; Andreas Brust; Norelle L Daly; Hartmut Cuny; David J Craik; Paul F Alewood; David J Adams; Richard J Lewis

doi:10.1016/j.bcp.2013.07.016

Isolation and characterization of α-conotoxin LsIA with potent activity at nicotinic acetylcholine receptors

Marco C Inserra, Shiva N Kompella, Irina Vetter, Andreas Brust, Norelle L Daly, Hartmut Cuny, David J Craik, Paul F Alewood, David J Adams, Richard J Lewis

Division of Cardiovascular Sciences (L5)

University of Queensland

Research output: Contribution to journal › Article › peer-review

Abstract

A new α-conotoxin LsIA was isolated from the crude venom of Conus limpusi using assay-guided RP-HPLC fractionation. Synthetic LsIA was a potent antagonist of α3β2, α3α5β2 and α7 nAChRs, with half-maximal inhibitory concentrations of 10, 31 and 10 nM, respectively. The structure of LsIA determined by NMR spectroscopy comprised a characteristic disulfide bond-stabilized α-helical structure and disordered N-terminal region. Potency reductions of up to 9-fold were observed for N-terminally truncated analogues of LsIA at α7 and α3β2 nAChRs, whereas C-terminal carboxylation enhanced potency 3-fold at α3β2 nAChRs but reduced potency 3-fold at α7 nAChRs. This study gives further insight into α-conotoxin pharmacology and the molecular basis of nAChR selectivity, highlighting the influence of N-terminal residues and C-terminal amidation on conotoxin pharmacology.

Original language	English
Pages (from-to)	791-799
Number of pages	9
Journal	Biochemical Pharmacology
Volume	86
Issue number	6
DOIs	https://doi.org/10.1016/j.bcp.2013.07.016
Publication status	Published - 15 Sept 2013

Keywords

Amino Acid Sequence
Animals
Calcium/metabolism
Cell Line, Tumor
Chromatography, Reverse-Phase
Conotoxins/chemical synthesis
Conus Snail/chemistry
Fluorescent Dyes
Humans
Magnetic Resonance Spectroscopy
Molecular Sequence Data
Nicotinic Antagonists/chemical synthesis
Protein Isoforms/antagonists & inhibitors
Protein Structure, Secondary
Quantitative Structure-Activity Relationship
Receptors, Nicotinic/metabolism
Substrate Specificity

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10.1016/j.bcp.2013.07.016

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title = "Isolation and characterization of α-conotoxin LsIA with potent activity at nicotinic acetylcholine receptors",

abstract = "A new α-conotoxin LsIA was isolated from the crude venom of Conus limpusi using assay-guided RP-HPLC fractionation. Synthetic LsIA was a potent antagonist of α3β2, α3α5β2 and α7 nAChRs, with half-maximal inhibitory concentrations of 10, 31 and 10 nM, respectively. The structure of LsIA determined by NMR spectroscopy comprised a characteristic disulfide bond-stabilized α-helical structure and disordered N-terminal region. Potency reductions of up to 9-fold were observed for N-terminally truncated analogues of LsIA at α7 and α3β2 nAChRs, whereas C-terminal carboxylation enhanced potency 3-fold at α3β2 nAChRs but reduced potency 3-fold at α7 nAChRs. This study gives further insight into α-conotoxin pharmacology and the molecular basis of nAChR selectivity, highlighting the influence of N-terminal residues and C-terminal amidation on conotoxin pharmacology.",

keywords = "Amino Acid Sequence, Animals, Calcium/metabolism, Cell Line, Tumor, Chromatography, Reverse-Phase, Conotoxins/chemical synthesis, Conus Snail/chemistry, Fluorescent Dyes, Humans, Magnetic Resonance Spectroscopy, Molecular Sequence Data, Nicotinic Antagonists/chemical synthesis, Protein Isoforms/antagonists & inhibitors, Protein Structure, Secondary, Quantitative Structure-Activity Relationship, Receptors, Nicotinic/metabolism, Substrate Specificity",

author = "Inserra, {Marco C} and Kompella, {Shiva N} and Irina Vetter and Andreas Brust and Daly, {Norelle L} and Hartmut Cuny and Craik, {David J} and Alewood, {Paul F} and Adams, {David J} and Lewis, {Richard J}",

note = "Copyright {\textcopyright} 2013. Published by Elsevier Inc.",

year = "2013",

month = sep,

day = "15",

doi = "10.1016/j.bcp.2013.07.016",

language = "English",

volume = "86",

pages = "791--799",

journal = "Biochemical Pharmacology",

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T1 - Isolation and characterization of α-conotoxin LsIA with potent activity at nicotinic acetylcholine receptors

AU - Inserra, Marco C

AU - Kompella, Shiva N

AU - Vetter, Irina

AU - Brust, Andreas

AU - Daly, Norelle L

AU - Cuny, Hartmut

AU - Craik, David J

AU - Alewood, Paul F

AU - Adams, David J

AU - Lewis, Richard J

PY - 2013/9/15

Y1 - 2013/9/15

N2 - A new α-conotoxin LsIA was isolated from the crude venom of Conus limpusi using assay-guided RP-HPLC fractionation. Synthetic LsIA was a potent antagonist of α3β2, α3α5β2 and α7 nAChRs, with half-maximal inhibitory concentrations of 10, 31 and 10 nM, respectively. The structure of LsIA determined by NMR spectroscopy comprised a characteristic disulfide bond-stabilized α-helical structure and disordered N-terminal region. Potency reductions of up to 9-fold were observed for N-terminally truncated analogues of LsIA at α7 and α3β2 nAChRs, whereas C-terminal carboxylation enhanced potency 3-fold at α3β2 nAChRs but reduced potency 3-fold at α7 nAChRs. This study gives further insight into α-conotoxin pharmacology and the molecular basis of nAChR selectivity, highlighting the influence of N-terminal residues and C-terminal amidation on conotoxin pharmacology.

AB - A new α-conotoxin LsIA was isolated from the crude venom of Conus limpusi using assay-guided RP-HPLC fractionation. Synthetic LsIA was a potent antagonist of α3β2, α3α5β2 and α7 nAChRs, with half-maximal inhibitory concentrations of 10, 31 and 10 nM, respectively. The structure of LsIA determined by NMR spectroscopy comprised a characteristic disulfide bond-stabilized α-helical structure and disordered N-terminal region. Potency reductions of up to 9-fold were observed for N-terminally truncated analogues of LsIA at α7 and α3β2 nAChRs, whereas C-terminal carboxylation enhanced potency 3-fold at α3β2 nAChRs but reduced potency 3-fold at α7 nAChRs. This study gives further insight into α-conotoxin pharmacology and the molecular basis of nAChR selectivity, highlighting the influence of N-terminal residues and C-terminal amidation on conotoxin pharmacology.

KW - Amino Acid Sequence

KW - Animals

KW - Calcium/metabolism

KW - Cell Line, Tumor

KW - Chromatography, Reverse-Phase

KW - Conotoxins/chemical synthesis

KW - Conus Snail/chemistry

KW - Fluorescent Dyes

KW - Humans

KW - Magnetic Resonance Spectroscopy

KW - Molecular Sequence Data

KW - Nicotinic Antagonists/chemical synthesis

KW - Protein Isoforms/antagonists & inhibitors

KW - Protein Structure, Secondary

KW - Quantitative Structure-Activity Relationship

KW - Receptors, Nicotinic/metabolism

KW - Substrate Specificity

U2 - 10.1016/j.bcp.2013.07.016

DO - 10.1016/j.bcp.2013.07.016

M3 - Article

C2 - 23924607

SN - 0006-2952

VL - 86

SP - 791

EP - 799

JO - Biochemical Pharmacology

JF - Biochemical Pharmacology

IS - 6

ER -

Isolation and characterization of α-conotoxin LsIA with potent activity at nicotinic acetylcholine receptors

Abstract

Keywords

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