Molecular determinants of subtype-selective efficacies of cytisine and the novel compound NS3861 at heteromeric nicotinic acetylcholine receptors

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Molecular determinants of subtype-selective efficacies of cytisine and the novel compound NS3861 at heteromeric nicotinic acetylcholine receptors. / Harpsøe, Kasper; Hald, Helle; Timmermann, Daniel B; Jensen, Marianne L; Dyhring, Tino; Nielsen, Elsebet Ø; Peters, Dan; Balle, Thomas; Gajhede, Michael; Kastrup, Jette Sandholm Jensen; Ahring, Philip K.

In: Journal of Biological Chemistry, Vol. 288, No. 4, 25.01.2013, p. 2559-2570.

Research output: Contribution to journalJournal articleResearchpeer-review

Harvard

Harpsøe, K, Hald, H, Timmermann, DB, Jensen, ML, Dyhring, T, Nielsen, EØ, Peters, D, Balle, T, Gajhede, M, Kastrup, JSJ & Ahring, PK 2013, 'Molecular determinants of subtype-selective efficacies of cytisine and the novel compound NS3861 at heteromeric nicotinic acetylcholine receptors', Journal of Biological Chemistry, vol. 288, no. 4, pp. 2559-2570. https://doi.org/10.1074/jbc.M112.436337

APA

Harpsøe, K., Hald, H., Timmermann, D. B., Jensen, M. L., Dyhring, T., Nielsen, E. Ø., ... Ahring, P. K. (2013). Molecular determinants of subtype-selective efficacies of cytisine and the novel compound NS3861 at heteromeric nicotinic acetylcholine receptors. Journal of Biological Chemistry, 288(4), 2559-2570. https://doi.org/10.1074/jbc.M112.436337

Vancouver

Harpsøe K, Hald H, Timmermann DB, Jensen ML, Dyhring T, Nielsen EØ et al. Molecular determinants of subtype-selective efficacies of cytisine and the novel compound NS3861 at heteromeric nicotinic acetylcholine receptors. Journal of Biological Chemistry. 2013 Jan 25;288(4):2559-2570. https://doi.org/10.1074/jbc.M112.436337

Author

Harpsøe, Kasper ; Hald, Helle ; Timmermann, Daniel B ; Jensen, Marianne L ; Dyhring, Tino ; Nielsen, Elsebet Ø ; Peters, Dan ; Balle, Thomas ; Gajhede, Michael ; Kastrup, Jette Sandholm Jensen ; Ahring, Philip K. / Molecular determinants of subtype-selective efficacies of cytisine and the novel compound NS3861 at heteromeric nicotinic acetylcholine receptors. In: Journal of Biological Chemistry. 2013 ; Vol. 288, No. 4. pp. 2559-2570.

Bibtex

@article{e7f078c199f24c7b9dff125ca08e5008,
title = "Molecular determinants of subtype-selective efficacies of cytisine and the novel compound NS3861 at heteromeric nicotinic acetylcholine receptors",
abstract = "Deciphering which specific agonist-receptor interactions affect efficacy levels is of high importance, because this will ultimately aid in designing selective drugs. The novel compound NS3861 and cytisine are agonists of nicotinic acetylcholine receptors (nAChRs) and both bind with high affinity to heteromeric a3{\ss}4 and a4{\ss}2 nAChRs. However, initial data revealed that the activation patterns of the two compounds show very distinct maximal efficacy readouts at various heteromeric nAChRs. To investigate the molecular determinants behind these observations, we performed in-depth patch clamp electrophysiological measurements of efficacy levels at heteromeric combinations of a3- and a4-, with {\ss}2- and {\ss}4-subunits, and various chimeric constructs thereof. Compared with cytisine, which selectively activates receptors containing {\ss}4- but not {\ss}2-subunits, NS3861 displays the opposite {\ss}-subunit preference and a complete lack of activation at a4-containing receptors. The maximal efficacy of NS3861 appeared solely dependent on the nature of the ligand-binding domain, whereas efficacy of cytisine was additionally affected by the nature of the {\ss}-subunit transmembrane domain. Molecular docking to nAChR subtype homology models suggests agonist specific interactions to two different residues on the complementary subunits as responsible for the {\ss}-subunit preference of both compounds. Furthermore, a principal subunit serine to threonine substitution may explain the lack of NS3861 activation at a4-containing receptors. In conclusion, our results are consistent with a hypothesis where agonist interactions with the principal subunit (a) primarily determine binding affinity, whereas interactions with key amino acids at the complementary subunit ({\ss}) affect agonist efficacy.",
author = "Kasper Harps{\o}e and Helle Hald and Timmermann, {Daniel B} and Jensen, {Marianne L} and Tino Dyhring and Nielsen, {Elsebet {\O}} and Dan Peters and Thomas Balle and Michael Gajhede and Kastrup, {Jette Sandholm Jensen} and Ahring, {Philip K}",
year = "2013",
month = "1",
day = "25",
doi = "10.1074/jbc.M112.436337",
language = "English",
volume = "288",
pages = "2559--2570",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology, Inc.",
number = "4",

}

RIS

TY - JOUR

T1 - Molecular determinants of subtype-selective efficacies of cytisine and the novel compound NS3861 at heteromeric nicotinic acetylcholine receptors

AU - Harpsøe, Kasper

AU - Hald, Helle

AU - Timmermann, Daniel B

AU - Jensen, Marianne L

AU - Dyhring, Tino

AU - Nielsen, Elsebet Ø

AU - Peters, Dan

AU - Balle, Thomas

AU - Gajhede, Michael

AU - Kastrup, Jette Sandholm Jensen

AU - Ahring, Philip K

PY - 2013/1/25

Y1 - 2013/1/25

N2 - Deciphering which specific agonist-receptor interactions affect efficacy levels is of high importance, because this will ultimately aid in designing selective drugs. The novel compound NS3861 and cytisine are agonists of nicotinic acetylcholine receptors (nAChRs) and both bind with high affinity to heteromeric a3ß4 and a4ß2 nAChRs. However, initial data revealed that the activation patterns of the two compounds show very distinct maximal efficacy readouts at various heteromeric nAChRs. To investigate the molecular determinants behind these observations, we performed in-depth patch clamp electrophysiological measurements of efficacy levels at heteromeric combinations of a3- and a4-, with ß2- and ß4-subunits, and various chimeric constructs thereof. Compared with cytisine, which selectively activates receptors containing ß4- but not ß2-subunits, NS3861 displays the opposite ß-subunit preference and a complete lack of activation at a4-containing receptors. The maximal efficacy of NS3861 appeared solely dependent on the nature of the ligand-binding domain, whereas efficacy of cytisine was additionally affected by the nature of the ß-subunit transmembrane domain. Molecular docking to nAChR subtype homology models suggests agonist specific interactions to two different residues on the complementary subunits as responsible for the ß-subunit preference of both compounds. Furthermore, a principal subunit serine to threonine substitution may explain the lack of NS3861 activation at a4-containing receptors. In conclusion, our results are consistent with a hypothesis where agonist interactions with the principal subunit (a) primarily determine binding affinity, whereas interactions with key amino acids at the complementary subunit (ß) affect agonist efficacy.

AB - Deciphering which specific agonist-receptor interactions affect efficacy levels is of high importance, because this will ultimately aid in designing selective drugs. The novel compound NS3861 and cytisine are agonists of nicotinic acetylcholine receptors (nAChRs) and both bind with high affinity to heteromeric a3ß4 and a4ß2 nAChRs. However, initial data revealed that the activation patterns of the two compounds show very distinct maximal efficacy readouts at various heteromeric nAChRs. To investigate the molecular determinants behind these observations, we performed in-depth patch clamp electrophysiological measurements of efficacy levels at heteromeric combinations of a3- and a4-, with ß2- and ß4-subunits, and various chimeric constructs thereof. Compared with cytisine, which selectively activates receptors containing ß4- but not ß2-subunits, NS3861 displays the opposite ß-subunit preference and a complete lack of activation at a4-containing receptors. The maximal efficacy of NS3861 appeared solely dependent on the nature of the ligand-binding domain, whereas efficacy of cytisine was additionally affected by the nature of the ß-subunit transmembrane domain. Molecular docking to nAChR subtype homology models suggests agonist specific interactions to two different residues on the complementary subunits as responsible for the ß-subunit preference of both compounds. Furthermore, a principal subunit serine to threonine substitution may explain the lack of NS3861 activation at a4-containing receptors. In conclusion, our results are consistent with a hypothesis where agonist interactions with the principal subunit (a) primarily determine binding affinity, whereas interactions with key amino acids at the complementary subunit (ß) affect agonist efficacy.

U2 - 10.1074/jbc.M112.436337

DO - 10.1074/jbc.M112.436337

M3 - Journal article

C2 - 23229547

VL - 288

SP - 2559

EP - 2570

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 4

ER -

ID: 44729105