Allosteric interactions with muscarinic acetylcholine receptors: Complex role of the conserved tryptophan M2422Trp in a critical cluster of amino acids for baseline affinity, subtype selectivity, and cooperativity

Stefanie Prilla, Jasmin Schrobang, John Ellis, Hans Dieter Höltje, Klaus Mohr

Research output: Contribution to journalArticle

52 Citations (Scopus)

Abstract

In general, the M2 subtype of muscarinic acetylcholine receptors has the highest sensitivity for allosteric modulators and the M5 subtype the lowest. The M2/M5 selectivity of some structurally diverse allosteric agents is known to be completely explained by M2177Tyr and M2423Thr in receptors whose orthosteric site is occupied by the conventional ligand N-methylscopolamine (NMS). This study explored the role of the conserved M 2422Trp and the adjacent M2423Thr in the binding of alkane-bisammonio type modulators, gallamine, and diallylcaracurine V. Experiments were performed with human M2 or M5 receptors or mutants thereof. It was found that M2 422Trp and M2423Thr independently influenced allosteric agent binding. The presence of M2423Thr may enhance the affinity of binding, depending on the allosteric agent, either directly or indirectly (by avoiding sterical hindrance through its M5 counterpart 478His). Replacement of M2422Trp and of the corresponding M5477Trp by alanine revealed a pronounced contribution of these epitopes to subtype independent baseline affinity in NMS-bound and NMS-free receptors for all agents except diallylcaracurine V. In a few instances, this tryptophan also influenced cooperativity and subtype selectivity. Docking simulations using a three-dimensional M2 receptor model revealed that the aromatic rings of M2177Tyr and M2422Trp, in a concerted action, might fix one of the aromatic moieties of alkane-bisammonio compounds between them. Thus, M2422Trp and the spatially adjacent M2177Tyr, as well as M2 423Thr, form a cluster of amino acids within the allosteric binding cleft that is pivotal for both M2/M5 subtype selectivity and baseline affinity of allosteric agents.

Original languageEnglish (US)
Pages (from-to)181-193
Number of pages13
JournalMolecular pharmacology
Volume70
Issue number1
DOIs
StatePublished - Jun 27 2006

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N-Methylscopolamine
Alkanes
Muscarinic Receptors
Tryptophan
Gallamine Triethiodide
Amino Acids
Alanine
Epitopes
Ligands

All Science Journal Classification (ASJC) codes

  • Molecular Medicine
  • Pharmacology

Cite this

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title = "Allosteric interactions with muscarinic acetylcholine receptors: Complex role of the conserved tryptophan M2422Trp in a critical cluster of amino acids for baseline affinity, subtype selectivity, and cooperativity",
abstract = "In general, the M2 subtype of muscarinic acetylcholine receptors has the highest sensitivity for allosteric modulators and the M5 subtype the lowest. The M2/M5 selectivity of some structurally diverse allosteric agents is known to be completely explained by M2177Tyr and M2423Thr in receptors whose orthosteric site is occupied by the conventional ligand N-methylscopolamine (NMS). This study explored the role of the conserved M 2422Trp and the adjacent M2423Thr in the binding of alkane-bisammonio type modulators, gallamine, and diallylcaracurine V. Experiments were performed with human M2 or M5 receptors or mutants thereof. It was found that M2 422Trp and M2423Thr independently influenced allosteric agent binding. The presence of M2423Thr may enhance the affinity of binding, depending on the allosteric agent, either directly or indirectly (by avoiding sterical hindrance through its M5 counterpart 478His). Replacement of M2422Trp and of the corresponding M5477Trp by alanine revealed a pronounced contribution of these epitopes to subtype independent baseline affinity in NMS-bound and NMS-free receptors for all agents except diallylcaracurine V. In a few instances, this tryptophan also influenced cooperativity and subtype selectivity. Docking simulations using a three-dimensional M2 receptor model revealed that the aromatic rings of M2177Tyr and M2422Trp, in a concerted action, might fix one of the aromatic moieties of alkane-bisammonio compounds between them. Thus, M2422Trp and the spatially adjacent M2177Tyr, as well as M2 423Thr, form a cluster of amino acids within the allosteric binding cleft that is pivotal for both M2/M5 subtype selectivity and baseline affinity of allosteric agents.",
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Allosteric interactions with muscarinic acetylcholine receptors : Complex role of the conserved tryptophan M2422Trp in a critical cluster of amino acids for baseline affinity, subtype selectivity, and cooperativity. / Prilla, Stefanie; Schrobang, Jasmin; Ellis, John; Höltje, Hans Dieter; Mohr, Klaus.

In: Molecular pharmacology, Vol. 70, No. 1, 27.06.2006, p. 181-193.

Research output: Contribution to journalArticle

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T2 - Complex role of the conserved tryptophan M2422Trp in a critical cluster of amino acids for baseline affinity, subtype selectivity, and cooperativity

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