Weak toxin from Naja kaouthia (WTX) belongs to the group of nonconventional "three-finger" snake neurotoxins. It irreversibly inhibits nicotinic acetylcholine receptors and allosterically interacts with muscarinic acetylcholine receptors (mAChRs). Using site-directed mutagenesis, NMR spectroscopy, and computer modeling, we investigated the recombinant mutant WTX analogue (rWTX) which, compared with the native toxin, has an additional N-terminal methionine residue. In comparison with the wild-type toxin, rWTX demonstrated an altered pharmacological profile, decreased binding of orthosteric antagonist N-methylscopolamine to human M1- and M2-mAChRs, and increased antagonist binding to M3-mAChR. Positively charged arginine residues located in the flexible loop II were found to be crucial for rWTX interactions with all types of mAChR. Computer modeling suggested that the rWTX loop II protrudes to the M1-mAChR allosteric ligand-binding site blocking the entrance to the orthosteric site. In contrast, toxin interacts with M3-mAChR by loop II without penetration into the allosteric site. Data obtained provide new structural insight into the target-specific allosteric regulation of mAChRs by "three-finger" snake neurotoxins.

• Klíčová slova
• G protein-coupled receptor (GPCR), computer modeling, nuclear magnetic resonance (NMR), protein dynamic, recombinant protein expression, site-directed mutagenesis, snake neurotoxin,
• MeSH
• Elapidae MeSH
• inzerční mutageneze MeSH
• jedy hadů čeledi Elapidae chemie   MeSH
• konformace proteinů MeSH
• molekulární sekvence - údaje MeSH
• neurotoxiny chemie   genetika   metabolismus   MeSH
• nukleární magnetická rezonance biomolekulární MeSH
• receptory muskarinové metabolismus   MeSH
• sekvence aminokyselin MeSH
• sekvenční homologie aminokyselin MeSH
• vazba proteinů MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• jedy hadů čeledi Elapidae MeSH
• neurotoxiny MeSH
• receptory muskarinové MeSH