Efficient screening of ligand-receptor complex formation using fluorescence labeling and size-exclusion chromatography.

D S Kulbatskii, M A Shulepko, N N Sluchanko, E O Yablokov, R A Kamyshinsky, Y M Chesnokov, M P Kirpichnikov, E N Lyukmanova

Biochemical and biophysical research communications 2020 Oct 29

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Evidence of a complex formation is a crucial step in the structural studies of ligand-receptor interactions. Here we presented a simple and fast approach for qualitative screening of the complex formation between the chimeric extracellular domain of the nicotinic acetylcholine receptor (α7-ECD) and three-finger proteins. Complex formation of snake toxins α-Bgtx and WTX, as well as of recombinant analogs of human proteins Lynx1 and SLURP-1, with α7-ECD was confirmed using fluorescently labeled ligands and size-exclusion chromatography with simultaneous absorbance and fluorescence detection. WTX/α7-ECD complex formation also was confirmed by cryo-EM. The proposed approach could easily be adopted to study the interaction of other receptors with their ligands. Copyright © 2020 Elsevier Inc. All rights reserved.

Citation

D S Kulbatskii, M A Shulepko, N N Sluchanko, E O Yablokov, R A Kamyshinsky, Y M Chesnokov, M P Kirpichnikov, E N Lyukmanova. Efficient screening of ligand-receptor complex formation using fluorescence labeling and size-exclusion chromatography. Biochemical and biophysical research communications. 2020 Oct 29;532(1):127-133