NMR structure and action on nicotinic acetylcholine receptors of water-soluble domain of human LYNX1.
Discovery of proteins expressed in the central nervous system sharing the three-finger structure with snake α-neurotoxins provoked much interest to their role in brain functions. Prototoxin LYNX1, having homology both to Ly6 proteins and three-finger neurotoxins, is the first identified member of this family membrane-tethered by a GPI anchor, which considerably complicates in vitro studies. We report for the first time the NMR spatial structure for the water-soluble domain of human LYNX1 lacking a GPI anchor (ws-LYNX1) and its concentration-dependent activity on nicotinic acetylcholine receptors (nAChRs). At 5-30 μM, ws-LYNX1 competed with (125)I-α-bungarotoxin for binding to the acetylcholine-binding proteins (AChBPs) and to Torpedo nAChR. Exposure of Xenopus oocytes expressing α7 nAChRs to 1 μM ws-LYNX1 enhanced the response to acetylcholine, but no effect was detected on α4β2 and α3β2 nAChRs. Increasing ws-LYNX1 concentration to 10 μM caused a modest inhibition of these three nAChR subtypes. A common feature for ws-LYNX1 and LYNX1 is a decrease of nAChR sensitivity to high concentrations of acetylcholine. NMR and functional analysis both demonstrate that ws-LYNX1 is an appropriate model to shed light on the mechanism of LYNX1 action. Computer modeling, based on ws-LYNX1 NMR structure and AChBP x-ray structure, revealed a possible mode of ws-LYNX1 binding.
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 16/10 Miklukho-Maklaya Street, 117997 Moscow, Russia.
Ekaterina N
EN
Lyukmanova
Zakhar O
ZO
Shenkarev
Mikhail A
MA
Shulepko
Konstantin S
KS
Mineev
Dieter
D
D'Hoedt
Igor E
IE
Kasheverov
Sergey Yu
SY
Filkin
Alexandra P
AP
Krivolapova
Helena
H
Janickova
Vladimir
V
Dolezal
Dmitry A
DA
Dolgikh
Alexander S
AS
Arseniev
Daniel
D
Bertrand
Victor I
VI
Tsetlin
Mikhail P
MP
Kirpichnikov
D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D002038:Bungarotoxins / Q000737:chemistry / Q000494:pharmacology; D058851:GPI-Linked Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular; D009865:Oocytes; D011485:Protein Binding; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary; D011978:Receptors, Nicotinic / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D012995:Solubility; D014982:Xenopus laevis
D048868:Adaptor Proteins, Signal Transducing; D002038:Bungarotoxins; D058851:GPI-Linked Proteins; C485626:LYNX1 protein, human; D011978:Receptors, Nicotinic
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