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Efficient screening of ligand-receptor complex formation using fluorescence labeling and size-exclusion chromatographyстатья
- Авторы: Kulbatskii D.S., Shulepko M.A., Sluchanko N.N., Yablokov E.O., Kamyshinsky R.A., Chesnokov Y.M., Kirpichnikov M.P., Lyukmanova E.N.
- Журнал: Biochemical and Biophysical Research Communications
- Год издания: 2020
- Издательство: Academic Press
- Местоположение издательства: United States
- DOI: 10.1016/j.bbrc.2020.08.021
- Аннотация: 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.
- Добавил в систему: Люкманова Екатерина Назымовна