| Autor: |
Prilla, Stefanie, Schrobang, Jasmin, Ellis, John, Höltje, Hans-Dieter, Mohr, Klaus |
| Zdroj: |
Molecular Pharmacology; July 2006, Vol. 70 Issue: 1 p181-193, 13p |
| Abstrakt: |
In general, the M2subtype of muscarinic acetylcholine receptors has the highest sensitivity for allosteric modulators and the M5subtype the lowest. The M2/M5selectivity 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 M2422Trp and the adjacent M2423Thr in the binding of alkane-bisammonio type modulators, gallamine, and diallylcaracurine V. Experiments were performed with human M2or M5receptors or mutants thereof. It was found that M2422Trp 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 M5counterpart 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 M2receptor 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 M2423Thr, form a cluster of amino acids within the allosteric binding cleft that is pivotal for both M2/M5subtype selectivity and baseline affinity of allosteric agents. |
| Databáze: |
Supplemental Index |
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