Crystal structure reveals canonical recognition of the phosphorylated cytoplasmic loop of human alpha7 nicotinic acetylcholine receptor by 14-3-3 protein

Crystal structure reveals canonical recognition of the phosphorylated cytoplasmic loop of human alpha7 nicotinic acetylcholine receptor by 14-3-3 protein Full article

Journal

Biochemical and Biophysical Research Communications
ISSN: 0006-291X , E-ISSN: 1090-2104

Output data

Year: 2023, Volume: 682, Pages: 91-96 Pages count : 6 DOI: 10.1016/j.bbrc.2023.09.086

Authors

Sluchanko Nikolai N. ¹ , Kapitonova Anna A. ¹ , Shulepko Mikhail A. ² , Kukushkin Ilya D. ^3,4 , Kulbatskii Dmitrii S. ⁴ , Tugaeva Kristina V. ¹ , Varfolomeeva Larisa A. ¹ , Minyaev Mikhail E. ⁵ , Boyko Konstantin M. ¹ , Popov Vladimir O. ^1,6 , Kirpichnikov Mikhail P. ^6,4 , Lyukmanova Ekaterina N. ^2,6,3,4

Affiliations

1	A.N. Bach Institute of Biochemistry, Federal Research Center of Biotechnology of the Russian Academy of Sciences, Moscow, 119071, Russia
2	Faculty of Biology, MSU-BIT Shenzhen University, Shenzhen, 518172, China
3	Phystech School of Biological and Medical Physics, Moscow Institute of Physics and Technology, Institutskiy per. 9, Dolgoprudny, Moscow region, 141701, Russia
4	Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences, Moscow, 117997, Russia
5	N.D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia
6	Interdisciplinary Scientific and Educational School of Moscow University “Molecular Technologies of the Living Systems and Synthetic Biology”, Biological Faculty, Lomonosov Moscow State University, Leninskie Gory, Moscow, 119234, Russia

Nicotinic acetylcholine receptors (nAChRs) are ligand-gated ion channels composed of five homologous subunits. The homopentameric α7-nAChR, abundantly expressed in the brain, is involved in the regulation of the neuronal plasticity and memory and undergoes phosphorylation by protein kinase A (PKA). Here, we extracted native α7-nAChR from murine brain, validated its assembly by cryo-EM and showed that phosphorylation by PKA in vitro enables its interaction with the abundant human brain protein 14-3-3ζ. Bioinformatic analysis narrowed the putative 14-3-3-binding site down to the fragment of the intracellular loop (ICL) containing Ser365 (Q361RRCSLASVEMS372), known to be phosphorylated in vivo. We reconstructed the 14-3-3ζ/ICL peptide complex and determined its structure by X-ray crystallography, which confirmed the Ser365 phosphorylation-dependent canonical recognition of the ICL by 14-3-3. A common mechanism of nAChRs’ regulation by ICL phosphorylation and 14-3-3 binding that potentially affects nAChR activity, stoichiometry, and surface expression is suggested.

Sluchanko N.N. , Kapitonova A.A. , Shulepko M.A. , Kukushkin I.D. , Kulbatskii D.S. , Tugaeva K.V. , Varfolomeeva L.A. , Minyaev M.E. , Boyko K.M. , Popov V.O. , Kirpichnikov M.P. , Lyukmanova E.N.
Crystal structure reveals canonical recognition of the phosphorylated cytoplasmic loop of human alpha7 nicotinic acetylcholine receptor by 14-3-3 protein
Biochemical and Biophysical Research Communications. 2023. V.682. P.91-96. DOI: 10.1016/j.bbrc.2023.09.086 WOS Scopus OpenAlex

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