Data from: Venom-gland transcriptome and venom proteome of the Malaysian king cobra (Ophiophagus hannah)

负责人：

关键词：

venom gland transcriptome;king cobra;venom proteome;nano-LCMS;ESI-LCMS\/MS;Ophiophagus hannah

DOI：

doi:10.5061/dryad.th547

摘要：

destruction in local envenoming. Dissimilarities in the subtypes and sequences between the neurotoxins of MOh and Naja kaouthia (monocled cobra) ar

Data from: Calcium imaging with genetically encoded sensor Case12: facile analysis of ?7/?9 nAChR mutants

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关键词：

Naja kaouthia;Xenopus laevis

DOI：

doi:10.5061/dryad.4k831

摘要：

Elucidation of the structural basis of pharmacological differences for highly homologous ?7 and ?9 nicotinic acetylcholine receptors (nAChRs) may shed light on their involvement in different physiological functions and diseases. Combination of site-directed mutagenesis and electrophysiology is a powerful tool to pinpoint the key amino-acid residues in the receptor ligand-binding site, but for ?7 and ?9 nAChRs it is complicated by their poor expression and fast desensitization. Here, we probed the ligand-binding properties of ?7/?9 nAChR mutants by a proposed simple and fast calcium imaging method. The method is based on transient co-expression of ?7/?9 nAChR mutants in neuroblastoma cells together with Ric-3 or NACHO chaperones and Case12 fluorescent calcium ion sensor followed by analysis of their pharmacology using a fluorescence microscope or a fluorometric imaging plate reader (FLIPR) with a GFP filter set. The results obtained were confirmed by electrophysiology and by calcium imaging with the conventional calcium indicator Fluo-4. The affinities for acetylcholine and epibatidine were determined for human and rat ?7 nAChRs, and for their mutants with homologous residues of ?9 nAChR incorporated at positions 117–119, 184, 185, 187, and 189, which are anticipated to be involved in ligand binding. The strongest decrease in the affinity was observed for mutations at positions 187 and 119. The L119D mutation of ?7 nAChR, showing a larger effect for epibatidine than for acetylcholine, may implicate this position in pharmacological differences between ?7 and ?9 nAChRs.