title: Secreted Isoform of Human Lynx1 (SLURP-2): Spatial Structure and Pharmacology of Interactions with Different Types of Acetylcholine Receptors.
abstract: Human-secreted Ly-6/uPAR-related protein-2 (SLURP-2) regulates the growth and differentiation of epithelial cells. Previously, the auto/paracrine activity of SLURP-2 was considered to be mediated via its interaction with the α3β2 subtype of the nicotinic acetylcholine receptors (nAChRs). Here, we describe the structure and pharmacology of a recombinant analogue of SLURP-2. Nuclear magnetic resonance spectroscopy revealed a 'three-finger' fold of SLURP-2 with a conserved β-structural core and three protruding loops. Affinity purification using cortical extracts revealed that SLURP-2 could interact with the α3, α4, α5, α7, β2, and β4 nAChR subunits, revealing its broader pharmacological profile. SLURP-2 inhibits acetylcholine-evoked currents at α4β2 and α3β2-nAChRs (IC50 ~0.17 and >3 μM, respectively) expressed in Xenopus oocytes. In contrast, at α7-nAChRs, SLURP-2 significantly enhances acetylcholine-evoked currents at concentrations <1 μM but induces inhibition at higher concentrations. SLURP-2 allosterically interacts with human M1 and M3 muscarinic acetylcholine receptors (mAChRs) that are overexpressed in CHO cells. SLURP-2 was found to promote the proliferation of human oral keratinocytes via interactions with α3β2-nAChRs, while it inhibited cell growth via α7-nAChRs. SLURP-2/mAChRs interactions are also probably involved in the control of keratinocyte growth. Computer modeling revealed possible SLURP-2 binding to the 'classical' orthosteric agonist/antagonist binding sites at α7 and α3β2-nAChRs.
authors.affiliation: Lomonosov Moscow State University, Leninskie Gori 1, Moscow 119234, Russian Federation.|Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry RAS, Miklukho-Maklaya Street 16/10, Moscow 117997, Russian Federation.
authors.forename: E N
authors.initials: EN
authors.lastname: Lyukmanova
authors.affiliation: Lomonosov Moscow State University, Leninskie Gori 1, Moscow 119234, Russian Federation.|Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry RAS, Miklukho-Maklaya Street 16/10, Moscow 117997, Russian Federation.
authors.forename: M A
authors.initials: MA
authors.lastname: Shulepko
authors.affiliation: Lomonosov Moscow State University, Leninskie Gori 1, Moscow 119234, Russian Federation.|Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry RAS, Miklukho-Maklaya Street 16/10, Moscow 117997, Russian Federation.|Moscow Institute of Physics and Technology, Institutskiy Pereulok 9, Dolgoprudny, Moscow Region 141700, Russian Federation.
authors.forename: Z O
authors.initials: ZO
authors.lastname: Shenkarev
authors.affiliation: Lomonosov Moscow State University, Leninskie Gori 1, Moscow 119234, Russian Federation.|Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry RAS, Miklukho-Maklaya Street 16/10, Moscow 117997, Russian Federation.
authors.forename: M L
authors.initials: ML
authors.lastname: Bychkov
authors.affiliation: Lomonosov Moscow State University, Leninskie Gori 1, Moscow 119234, Russian Federation.|Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry RAS, Miklukho-Maklaya Street 16/10, Moscow 117997, Russian Federation.
authors.forename: A S
authors.initials: AS
authors.lastname: Paramonov
authors.affiliation: Lomonosov Moscow State University, Leninskie Gori 1, Moscow 119234, Russian Federation.|Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry RAS, Miklukho-Maklaya Street 16/10, Moscow 117997, Russian Federation.
authors.forename: A O
authors.initials: AO
authors.lastname: Chugunov
authors.affiliation: Lomonosov Moscow State University, Leninskie Gori 1, Moscow 119234, Russian Federation.|Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry RAS, Miklukho-Maklaya Street 16/10, Moscow 117997, Russian Federation.
authors.forename: D S
authors.initials: DS
authors.lastname: Kulbatskii
authors.affiliation: Department of Drug Design and Pharmacology, University of Copenhagen, Jagtvej 160, DK-2100 Copenhagen, Denmark.
authors.forename: M
authors.initials: M
authors.lastname: Arvaniti
authors.affiliation: Institute of Physiology, Academy of Sciences of the Czech Republic (public research institution), Prague, 14220, Czech Republic.
authors.forename: Eva
authors.initials: E
authors.lastname: Dolejsi
authors.affiliation: HiQScreen Sàrl, 6 rte de Compois, 1222, Vésenaz, Geneva, Switzerland.
authors.forename: T
authors.initials: T
authors.lastname: Schaer
authors.affiliation: Lomonosov Moscow State University, Leninskie Gori 1, Moscow 119234, Russian Federation.|Moscow Institute of Physics and Technology, Institutskiy Pereulok 9, Dolgoprudny, Moscow Region 141700, Russian Federation.
authors.forename: A S
authors.initials: AS
authors.lastname: Arseniev
authors.affiliation: Lomonosov Moscow State University, Leninskie Gori 1, Moscow 119234, Russian Federation.|National Research University Higher School of Economics, Myasnitskaya ul. 20, 101000 Moscow, Russia.
authors.forename: R G
authors.initials: RG
authors.lastname: Efremov
authors.affiliation: Department of Drug Design and Pharmacology, University of Copenhagen, Jagtvej 160, DK-2100 Copenhagen, Denmark.
authors.forename: M S
authors.initials: MS
authors.lastname: Thomsen
authors.affiliation: Institute of Physiology, Academy of Sciences of the Czech Republic (public research institution), Prague, 14220, Czech Republic.
authors.forename: V
authors.initials: V
authors.lastname: Dolezal
authors.affiliation: HiQScreen Sàrl, 6 rte de Compois, 1222, Vésenaz, Geneva, Switzerland.
authors.forename: D
authors.initials: D
authors.lastname: Bertrand
authors.affiliation: Lomonosov Moscow State University, Leninskie Gori 1, Moscow 119234, Russian Federation.|Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry RAS, Miklukho-Maklaya Street 16/10, Moscow 117997, Russian Federation.
authors.forename: D A
authors.initials: DA
authors.lastname: Dolgikh
authors.affiliation: Lomonosov Moscow State University, Leninskie Gori 1, Moscow 119234, Russian Federation.|Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry RAS, Miklukho-Maklaya Street 16/10, Moscow 117997, Russian Federation.
authors.forename: M P
authors.initials: MP
authors.lastname: Kirpichnikov
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D000328:Adult; D000818:Animals; D001665:Binding Sites / Q000502:physiology; D016466:CHO Cells; D002460:Cell Line; D049109:Cell Proliferation / Q000502:physiology; D003198:Computer Simulation; D003412:Cricetulus; D004833:Epilepsy, Temporal Lobe / Q000473:pathology; D005071:Evoked Potentials / Q000502:physiology*; D005260:Female; D058851:GPI-Linked Proteins / Q000378:metabolism*; D006801:Humans; D015603:Keratinocytes / Q000378:metabolism*; D008875:Middle Aged; D019906:Nuclear Magnetic Resonance, Biomolecular; D009865:Oocytes / Q000378:metabolism; D016716:PC12 Cells; D011485:Protein Binding / Q000502:physiology; D051381:Rats; D011976:Receptors, Muscarinic / Q000378:metabolism*; D011978:Receptors, Nicotinic / Q000378:metabolism*; D014981:Xenopus; D064569:alpha7 Nicotinic Acetylcholine Receptor / Q000378:metabolism*
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D058851:GPI-Linked Proteins; C485626:LYNX1 protein, human; D011976:Receptors, Muscarinic; D011978:Receptors, Nicotinic; D064569:alpha7 Nicotinic Acetylcholine Receptor; C419362:nicotinic receptor alpha3beta2