Human Three-Finger Protein Lypd6 Is a Negative Modulator of the Cholinergic System in the Brain
Lypd6 is a GPI-tethered protein from the Ly-6/uPAR family expressed in the brain. Lypd6 enhances the Wnt/β-catenin signaling, although its action on nicotinic acetylcholine receptors (nAChRs) have been also proposed. To investigate a cholinergic activity of Lypd6, we studied a recombinant water-solu...
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Frontiers Media S.A.
2021-09-01
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Online Access: | https://www.frontiersin.org/articles/10.3389/fcell.2021.662227/full |
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Article |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Dmitrii Kulbatskii Zakhar Shenkarev Zakhar Shenkarev Maxim Bychkov Eugene Loktyushov Mikhail Shulepko Sergey Koshelev Igor Povarov Alexander Popov Alexander Popov Steve Peigneur Anton Chugunov Anton Chugunov Anton Chugunov Sergey Kozlov Irina Sharonova Roman Efremov Roman Efremov Roman Efremov Vladimir Skrebitsky Jan Tytgat Mikhail Kirpichnikov Mikhail Kirpichnikov Ekaterina Lyukmanova Ekaterina Lyukmanova Ekaterina Lyukmanova |
spellingShingle |
Dmitrii Kulbatskii Zakhar Shenkarev Zakhar Shenkarev Maxim Bychkov Eugene Loktyushov Mikhail Shulepko Sergey Koshelev Igor Povarov Alexander Popov Alexander Popov Steve Peigneur Anton Chugunov Anton Chugunov Anton Chugunov Sergey Kozlov Irina Sharonova Roman Efremov Roman Efremov Roman Efremov Vladimir Skrebitsky Jan Tytgat Mikhail Kirpichnikov Mikhail Kirpichnikov Ekaterina Lyukmanova Ekaterina Lyukmanova Ekaterina Lyukmanova Human Three-Finger Protein Lypd6 Is a Negative Modulator of the Cholinergic System in the Brain Frontiers in Cell and Developmental Biology Lypd6 nAChR cognitive function synaptic plasticity Ly6/uPAR three-finger |
author_facet |
Dmitrii Kulbatskii Zakhar Shenkarev Zakhar Shenkarev Maxim Bychkov Eugene Loktyushov Mikhail Shulepko Sergey Koshelev Igor Povarov Alexander Popov Alexander Popov Steve Peigneur Anton Chugunov Anton Chugunov Anton Chugunov Sergey Kozlov Irina Sharonova Roman Efremov Roman Efremov Roman Efremov Vladimir Skrebitsky Jan Tytgat Mikhail Kirpichnikov Mikhail Kirpichnikov Ekaterina Lyukmanova Ekaterina Lyukmanova Ekaterina Lyukmanova |
author_sort |
Dmitrii Kulbatskii |
title |
Human Three-Finger Protein Lypd6 Is a Negative Modulator of the Cholinergic System in the Brain |
title_short |
Human Three-Finger Protein Lypd6 Is a Negative Modulator of the Cholinergic System in the Brain |
title_full |
Human Three-Finger Protein Lypd6 Is a Negative Modulator of the Cholinergic System in the Brain |
title_fullStr |
Human Three-Finger Protein Lypd6 Is a Negative Modulator of the Cholinergic System in the Brain |
title_full_unstemmed |
Human Three-Finger Protein Lypd6 Is a Negative Modulator of the Cholinergic System in the Brain |
title_sort |
human three-finger protein lypd6 is a negative modulator of the cholinergic system in the brain |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Cell and Developmental Biology |
issn |
2296-634X |
publishDate |
2021-09-01 |
description |
Lypd6 is a GPI-tethered protein from the Ly-6/uPAR family expressed in the brain. Lypd6 enhances the Wnt/β-catenin signaling, although its action on nicotinic acetylcholine receptors (nAChRs) have been also proposed. To investigate a cholinergic activity of Lypd6, we studied a recombinant water-soluble variant of the human protein (ws-Lypd6) containing isolated “three-finger” LU-domain. Experiments at different nAChR subtypes expressed in Xenopus oocytes revealed the negative allosteric modulatory activity of ws-Lypd6. Ws-Lypd6 inhibited ACh-evoked currents at α3β4- and α7-nAChRs with IC50 of ∼35 and 10 μM, respectively, and the maximal amplitude of inhibition of 30–50%. EC50 of ACh at α3β4-nAChRs (∼30 μM) was not changed in the presence of 35 μM ws-Lypd6, while the maximal amplitude of ACh-evoked current was reduced by ∼20%. Ws-Lypd6 did not elicit currents through nAChRs in the absence of ACh. Application of 1 μM ws-Lypd6 significantly inhibited (up to ∼28%) choline-evoked current at α7-nAChRs in rat hippocampal slices. Similar to snake neurotoxin α-bungarotoxin, ws-Lypd6 suppressed the long-term potentiation (LTP) in mouse hippocampal slices. Colocalization of endogenous GPI-tethered Lypd6 with α3β4- and α7-nAChRs was detected in primary cortical and hippocampal neurons. Ws-Lypd6 interaction with the extracellular domain of α7-nAChR was modeled using the ensemble protein-protein docking protocol. The interaction of all three Lypd6 loops (“fingers”) with the entrance to the orthosteric ligand-binding site and the loop C of the primary receptor subunit was predicted. The results obtained allow us to consider Lypd6 as the endogenous negative modulator involved in the regulation of the cholinergic system in the brain. |
topic |
Lypd6 nAChR cognitive function synaptic plasticity Ly6/uPAR three-finger |
url |
https://www.frontiersin.org/articles/10.3389/fcell.2021.662227/full |
work_keys_str_mv |
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doaj-a865cba71d844068a019acd3c7b696bb2021-09-22T15:52:28ZengFrontiers Media S.A.Frontiers in Cell and Developmental Biology2296-634X2021-09-01910.3389/fcell.2021.662227662227Human Three-Finger Protein Lypd6 Is a Negative Modulator of the Cholinergic System in the BrainDmitrii Kulbatskii0Zakhar Shenkarev1Zakhar Shenkarev2Maxim Bychkov3Eugene Loktyushov4Mikhail Shulepko5Sergey Koshelev6Igor Povarov7Alexander Popov8Alexander Popov9Steve Peigneur10Anton Chugunov11Anton Chugunov12Anton Chugunov13Sergey Kozlov14Irina Sharonova15Roman Efremov16Roman Efremov17Roman Efremov18Vladimir Skrebitsky19Jan Tytgat20Mikhail Kirpichnikov21Mikhail Kirpichnikov22Ekaterina Lyukmanova23Ekaterina Lyukmanova24Ekaterina Lyukmanova25Bioengineering Department, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry Russian Academy of Sciences (RAS), Moscow, RussiaStructural Biology Department, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry Russian Academy of Sciences (RAS), Moscow, RussiaPhystech School of Biological and Medical Physics, Moscow Institute of Physics and Technology, Moscow, RussiaBioengineering Department, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry Russian Academy of Sciences (RAS), Moscow, RussiaBioengineering Department, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry Russian Academy of Sciences (RAS), Moscow, RussiaBioengineering Department, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry Russian Academy of Sciences (RAS), Moscow, RussiaDepartment of Molecular Neurobiology, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry Russian Academy of Sciences (RAS), Moscow, RussiaBrain Research Department, Research Center of Neurology, Moscow, RussiaDepartment of Molecular Neurobiology, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry Russian Academy of Sciences (RAS), Moscow, RussiaInstitute of Neuroscience, Nizhny Novgorod University, Nizhny Novgorod, RussiaToxicology and Pharmacology, University of Leuven (KU Leuven), Leuven, BelgiumStructural Biology Department, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry Russian Academy of Sciences (RAS), Moscow, RussiaPhystech School of Biological and Medical Physics, Moscow Institute of Physics and Technology, Moscow, RussiaInternational Laboratory for Supercomputer Atomistic Modelling and Multi-Scale Analysis, National Research University Higher School of Economics, Moscow, RussiaDepartment of Molecular Neurobiology, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry Russian Academy of Sciences (RAS), Moscow, RussiaBrain Research Department, Research Center of Neurology, Moscow, RussiaStructural Biology Department, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry Russian Academy of Sciences (RAS), Moscow, RussiaPhystech School of Biological and Medical Physics, Moscow Institute of Physics and Technology, Moscow, RussiaInternational Laboratory for Supercomputer Atomistic Modelling and Multi-Scale Analysis, National Research University Higher School of Economics, Moscow, RussiaBrain Research Department, Research Center of Neurology, Moscow, RussiaToxicology and Pharmacology, University of Leuven (KU Leuven), Leuven, BelgiumBioengineering Department, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry Russian Academy of Sciences (RAS), Moscow, RussiaBiological Faculty, Lomonosov Moscow State University, Moscow, RussiaBioengineering Department, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry Russian Academy of Sciences (RAS), Moscow, RussiaPhystech School of Biological and Medical Physics, Moscow Institute of Physics and Technology, Moscow, RussiaBiological Faculty, Lomonosov Moscow State University, Moscow, RussiaLypd6 is a GPI-tethered protein from the Ly-6/uPAR family expressed in the brain. Lypd6 enhances the Wnt/β-catenin signaling, although its action on nicotinic acetylcholine receptors (nAChRs) have been also proposed. To investigate a cholinergic activity of Lypd6, we studied a recombinant water-soluble variant of the human protein (ws-Lypd6) containing isolated “three-finger” LU-domain. Experiments at different nAChR subtypes expressed in Xenopus oocytes revealed the negative allosteric modulatory activity of ws-Lypd6. Ws-Lypd6 inhibited ACh-evoked currents at α3β4- and α7-nAChRs with IC50 of ∼35 and 10 μM, respectively, and the maximal amplitude of inhibition of 30–50%. EC50 of ACh at α3β4-nAChRs (∼30 μM) was not changed in the presence of 35 μM ws-Lypd6, while the maximal amplitude of ACh-evoked current was reduced by ∼20%. Ws-Lypd6 did not elicit currents through nAChRs in the absence of ACh. Application of 1 μM ws-Lypd6 significantly inhibited (up to ∼28%) choline-evoked current at α7-nAChRs in rat hippocampal slices. Similar to snake neurotoxin α-bungarotoxin, ws-Lypd6 suppressed the long-term potentiation (LTP) in mouse hippocampal slices. Colocalization of endogenous GPI-tethered Lypd6 with α3β4- and α7-nAChRs was detected in primary cortical and hippocampal neurons. Ws-Lypd6 interaction with the extracellular domain of α7-nAChR was modeled using the ensemble protein-protein docking protocol. The interaction of all three Lypd6 loops (“fingers”) with the entrance to the orthosteric ligand-binding site and the loop C of the primary receptor subunit was predicted. The results obtained allow us to consider Lypd6 as the endogenous negative modulator involved in the regulation of the cholinergic system in the brain.https://www.frontiersin.org/articles/10.3389/fcell.2021.662227/fullLypd6nAChRcognitive functionsynaptic plasticityLy6/uPARthree-finger |