Generation of candidate ligands for nicotinic acetylcholine receptors via in situ click chemistry with a soluble acetylcholine binding protein template

Neil P. Grimster, Bernhard Stump, Joseph R. Fotsing, Timo Weide, Todd T. Talley, John G. Yamauchi, Ákos Nemecz, Choel Kim, Kwok Yiu Ho, K. Barry Sharpless, Palmer Taylor, Valery V. Fokin

Research output: Contribution to journalArticle

49 Citations (Scopus)

Abstract

Nicotinic acetylcholine receptors (nAChRs), which are responsible for mediating key physiological functions, are ubiquitous in the central and peripheral nervous systems. As members of the Cys loop ligand-gated ion channel family, neuronal nAChRs are pentameric, composed of various permutations of α (α2 to α10) and β (β2 to β4) subunits forming functional heteromeric or homomeric receptors. Diversity in nAChR subunit composition complicates the development of selective ligands for specific subtypes, since the five binding sites reside at the subunit interfaces. The acetylcholine binding protein (AChBP), a soluble extracellular domain homologue secreted by mollusks, serves as a general structural surrogate for the nAChRs. In this work, homomeric AChBPs from Lymnaea and Aplysia snails were used as in situ templates for the generation of novel and potent ligands that selectively bind to these proteins. The cycloaddition reaction between building-block azides and alkynes to form stable 1,2,3-triazoles was used to generate the leads. The extent of triazole formation on the AChBP template correlated with the affinity of the triazole product for the nicotinic ligand binding site. Instead of the in situ protein-templated azide-alkyne cycloaddition reaction occurring at a localized, sequestered enzyme active center as previously shown, we demonstrate that the in situ reaction can take place at the subunit interfaces of an oligomeric protein and can thus be used as a tool for identifying novel candidate nAChR ligands. The crystal structure of one of the in situ-formed triazole-AChBP complexes shows binding poses and molecular determinants of interactions predicted from structures of known agonists and antagonists. Hence, the click chemistry approach with an in situ template of a receptor provides a novel synthetic avenue for generating candidate agonists and antagonists for ligand-gated ion channels.

Original languageEnglish
Pages (from-to)6732-6740
Number of pages9
JournalJournal of the American Chemical Society
Volume134
Issue number15
DOIs
Publication statusPublished - 18 Apr 2012
Externally publishedYes

Fingerprint

Click Chemistry
Triazoles
Nicotinic Receptors
Acetylcholine
Carrier Proteins
Ligands
Ligand-Gated Ion Channels
Alkynes
Azides
Cycloaddition
Cycloaddition Reaction
Binding Sites
Binding sites
Lymnaea
Proteins
Aplysia
Mollusca
Snails
Peripheral Nervous System
Neurology

ASJC Scopus subject areas

  • Chemistry(all)
  • Catalysis
  • Biochemistry
  • Colloid and Surface Chemistry

Cite this

Generation of candidate ligands for nicotinic acetylcholine receptors via in situ click chemistry with a soluble acetylcholine binding protein template. / Grimster, Neil P.; Stump, Bernhard; Fotsing, Joseph R.; Weide, Timo; Talley, Todd T.; Yamauchi, John G.; Nemecz, Ákos; Kim, Choel; Ho, Kwok Yiu; Sharpless, K. Barry; Taylor, Palmer; Fokin, Valery V.

In: Journal of the American Chemical Society, Vol. 134, No. 15, 18.04.2012, p. 6732-6740.

Research output: Contribution to journalArticle

Grimster, NP, Stump, B, Fotsing, JR, Weide, T, Talley, TT, Yamauchi, JG, Nemecz, Á, Kim, C, Ho, KY, Sharpless, KB, Taylor, P & Fokin, VV 2012, 'Generation of candidate ligands for nicotinic acetylcholine receptors via in situ click chemistry with a soluble acetylcholine binding protein template', Journal of the American Chemical Society, vol. 134, no. 15, pp. 6732-6740. https://doi.org/10.1021/ja3001858
Grimster, Neil P. ; Stump, Bernhard ; Fotsing, Joseph R. ; Weide, Timo ; Talley, Todd T. ; Yamauchi, John G. ; Nemecz, Ákos ; Kim, Choel ; Ho, Kwok Yiu ; Sharpless, K. Barry ; Taylor, Palmer ; Fokin, Valery V. / Generation of candidate ligands for nicotinic acetylcholine receptors via in situ click chemistry with a soluble acetylcholine binding protein template. In: Journal of the American Chemical Society. 2012 ; Vol. 134, No. 15. pp. 6732-6740.
@article{016783aeb6e041c9b2e777f5af11532f,
title = "Generation of candidate ligands for nicotinic acetylcholine receptors via in situ click chemistry with a soluble acetylcholine binding protein template",
abstract = "Nicotinic acetylcholine receptors (nAChRs), which are responsible for mediating key physiological functions, are ubiquitous in the central and peripheral nervous systems. As members of the Cys loop ligand-gated ion channel family, neuronal nAChRs are pentameric, composed of various permutations of α (α2 to α10) and β (β2 to β4) subunits forming functional heteromeric or homomeric receptors. Diversity in nAChR subunit composition complicates the development of selective ligands for specific subtypes, since the five binding sites reside at the subunit interfaces. The acetylcholine binding protein (AChBP), a soluble extracellular domain homologue secreted by mollusks, serves as a general structural surrogate for the nAChRs. In this work, homomeric AChBPs from Lymnaea and Aplysia snails were used as in situ templates for the generation of novel and potent ligands that selectively bind to these proteins. The cycloaddition reaction between building-block azides and alkynes to form stable 1,2,3-triazoles was used to generate the leads. The extent of triazole formation on the AChBP template correlated with the affinity of the triazole product for the nicotinic ligand binding site. Instead of the in situ protein-templated azide-alkyne cycloaddition reaction occurring at a localized, sequestered enzyme active center as previously shown, we demonstrate that the in situ reaction can take place at the subunit interfaces of an oligomeric protein and can thus be used as a tool for identifying novel candidate nAChR ligands. The crystal structure of one of the in situ-formed triazole-AChBP complexes shows binding poses and molecular determinants of interactions predicted from structures of known agonists and antagonists. Hence, the click chemistry approach with an in situ template of a receptor provides a novel synthetic avenue for generating candidate agonists and antagonists for ligand-gated ion channels.",
author = "Grimster, {Neil P.} and Bernhard Stump and Fotsing, {Joseph R.} and Timo Weide and Talley, {Todd T.} and Yamauchi, {John G.} and {\'A}kos Nemecz and Choel Kim and Ho, {Kwok Yiu} and Sharpless, {K. Barry} and Palmer Taylor and Fokin, {Valery V.}",
year = "2012",
month = "4",
day = "18",
doi = "10.1021/ja3001858",
language = "English",
volume = "134",
pages = "6732--6740",
journal = "Journal of the American Chemical Society",
issn = "0002-7863",
publisher = "American Chemical Society",
number = "15",

}

TY - JOUR

T1 - Generation of candidate ligands for nicotinic acetylcholine receptors via in situ click chemistry with a soluble acetylcholine binding protein template

AU - Grimster, Neil P.

AU - Stump, Bernhard

AU - Fotsing, Joseph R.

AU - Weide, Timo

AU - Talley, Todd T.

AU - Yamauchi, John G.

AU - Nemecz, Ákos

AU - Kim, Choel

AU - Ho, Kwok Yiu

AU - Sharpless, K. Barry

AU - Taylor, Palmer

AU - Fokin, Valery V.

PY - 2012/4/18

Y1 - 2012/4/18

N2 - Nicotinic acetylcholine receptors (nAChRs), which are responsible for mediating key physiological functions, are ubiquitous in the central and peripheral nervous systems. As members of the Cys loop ligand-gated ion channel family, neuronal nAChRs are pentameric, composed of various permutations of α (α2 to α10) and β (β2 to β4) subunits forming functional heteromeric or homomeric receptors. Diversity in nAChR subunit composition complicates the development of selective ligands for specific subtypes, since the five binding sites reside at the subunit interfaces. The acetylcholine binding protein (AChBP), a soluble extracellular domain homologue secreted by mollusks, serves as a general structural surrogate for the nAChRs. In this work, homomeric AChBPs from Lymnaea and Aplysia snails were used as in situ templates for the generation of novel and potent ligands that selectively bind to these proteins. The cycloaddition reaction between building-block azides and alkynes to form stable 1,2,3-triazoles was used to generate the leads. The extent of triazole formation on the AChBP template correlated with the affinity of the triazole product for the nicotinic ligand binding site. Instead of the in situ protein-templated azide-alkyne cycloaddition reaction occurring at a localized, sequestered enzyme active center as previously shown, we demonstrate that the in situ reaction can take place at the subunit interfaces of an oligomeric protein and can thus be used as a tool for identifying novel candidate nAChR ligands. The crystal structure of one of the in situ-formed triazole-AChBP complexes shows binding poses and molecular determinants of interactions predicted from structures of known agonists and antagonists. Hence, the click chemistry approach with an in situ template of a receptor provides a novel synthetic avenue for generating candidate agonists and antagonists for ligand-gated ion channels.

AB - Nicotinic acetylcholine receptors (nAChRs), which are responsible for mediating key physiological functions, are ubiquitous in the central and peripheral nervous systems. As members of the Cys loop ligand-gated ion channel family, neuronal nAChRs are pentameric, composed of various permutations of α (α2 to α10) and β (β2 to β4) subunits forming functional heteromeric or homomeric receptors. Diversity in nAChR subunit composition complicates the development of selective ligands for specific subtypes, since the five binding sites reside at the subunit interfaces. The acetylcholine binding protein (AChBP), a soluble extracellular domain homologue secreted by mollusks, serves as a general structural surrogate for the nAChRs. In this work, homomeric AChBPs from Lymnaea and Aplysia snails were used as in situ templates for the generation of novel and potent ligands that selectively bind to these proteins. The cycloaddition reaction between building-block azides and alkynes to form stable 1,2,3-triazoles was used to generate the leads. The extent of triazole formation on the AChBP template correlated with the affinity of the triazole product for the nicotinic ligand binding site. Instead of the in situ protein-templated azide-alkyne cycloaddition reaction occurring at a localized, sequestered enzyme active center as previously shown, we demonstrate that the in situ reaction can take place at the subunit interfaces of an oligomeric protein and can thus be used as a tool for identifying novel candidate nAChR ligands. The crystal structure of one of the in situ-formed triazole-AChBP complexes shows binding poses and molecular determinants of interactions predicted from structures of known agonists and antagonists. Hence, the click chemistry approach with an in situ template of a receptor provides a novel synthetic avenue for generating candidate agonists and antagonists for ligand-gated ion channels.

UR - http://www.scopus.com/inward/record.url?scp=84859994538&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84859994538&partnerID=8YFLogxK

U2 - 10.1021/ja3001858

DO - 10.1021/ja3001858

M3 - Article

VL - 134

SP - 6732

EP - 6740

JO - Journal of the American Chemical Society

JF - Journal of the American Chemical Society

SN - 0002-7863

IS - 15

ER -