Nicotinic receptor pharmacology in silico: Insights and challenges

Gulsevin, Alican

doi:10.1016/j.neuropharm.2020.108257

Cited by 8 publications

(7 citation statements)

References 151 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Loop I was proposed to interact with the loop C of the primary receptor’s subunit. The loop C covers the orthosteric ligand binding site of the nicotinic receptor, and its movement is coupled with the receptor activation ( Law et al, 2005 ; Gulsevin, 2020 ). Loop II and loop III of SLURP-1 were supposed to form additional contacts with the primary and complementary receptor subunits ( Shulepko et al, 2021 ).…”

Section: Discussionmentioning

confidence: 99%

SLURP-1 Controls Growth and Migration of Lung Adenocarcinoma Cells, Forming a Complex With α7-nAChR and PDGFR/EGFR Heterodimer

Bychkov

Shulepko

Shlepova

et al. 2021

Front. Cell Dev. Biol.

View full text Add to dashboard Cite

Secreted Ly6/uPAR-related protein 1 (SLURP-1) is a secreted Ly6/uPAR protein that negatively modulates the nicotinic acetylcholine receptor of α7 type (α7-nAChR), participating in control of cancer cell growth. Previously we showed, that a recombinant analogue of human SLURP-1 (rSLURP-1) diminishes the lung adenocarcinoma A549 cell proliferation and abolishes the nicotine-induced growth stimulation. Here, using multiplex immunoassay, we demonstrated a decrease in PTEN and mammalian target of rapamycin (mTOR) kinase phosphorylation in A549 cells upon the rSLURP-1 treatment pointing on down-regulation of the PI3K/AKT/mTOR signaling pathway. Decreased phosphorylation of the platelet-derived growth factor receptor type β (PDGFRβ) and arrest of the A549 cell cycle in the S and G2/M phases without apoptosis induction was also observed. Using a scratch migration assay, inhibition of A549 cell migration under the rSLURP-1 treatment was found. Affinity extraction demonstrated that rSLURP-1 in A549 cells forms a complex not only with α7-nAChR, but also with PDGFRα and epidermal growth factor receptor (EGFR), which are known to be involved in regulation of cancer cell growth and migration and are able to form a heterodimer. Knock-down of the genes encoding α7-nAChR, PDGFRα, and EGFR confirmed the involvement of these receptors in the anti-migration effect of SLURP-1. Thus, SLURP-1 can target the α7-nAChR complexes with PDGFRα and EGFR in the membrane of epithelial cells. Using chimeric proteins with grafted SLURP-1 loops we demonstrated that loop I is the principal active site responsible for the SLURP-1 interaction with α7-nAChR and its antiproliferative effect. Synthetic peptide mimicking the loop I cyclized by a disulfide bond inhibited ACh-evoked current at α7-nAChR, as well as A549 cell proliferation and migration. This synthetic peptide represents a promising prototype of new antitumor drug with the properties close to that of the native SLURP-1 protein.

show abstract

Section: Discussionmentioning

confidence: 99%

SLURP-1 Controls Growth and Migration of Lung Adenocarcinoma Cells, Forming a Complex With α7-nAChR and PDGFR/EGFR Heterodimer

Bychkov

Shulepko

Shlepova

et al. 2021

Front. Cell Dev. Biol.

View full text Add to dashboard Cite

show abstract

“…The “open” position of the loop C is usually observed in the complexes with the antagonists (e.g., α-Bgtx, conotoxin ImI) and corresponds to the closed channel pore, while the “closed” ligand-binding pocket was found in the complexes with the agonists (e.g., epibatidine, lobeline, nicotine) ( Hansen et al, 2005 ). The movement of the loop C (its closure) triggers the conformational changes that lead to the opening of the channel pore ( Law et al, 2005 ; Gulsevin, 2020 ; Noviello et al, 2021 ). The obtained docking solution ( Figures 8B,C ) indicates that the loops I, II, and III of ws-Lypd6 may interact with α7-nAChRs at the entrance to the orthosteric ligand-binding pocket.…”

Section: Discussionmentioning

confidence: 99%

Human Three-Finger Protein Lypd6 Is a Negative Modulator of the Cholinergic System in the Brain

Kulbatskii

Шенкарев

Bychkov

et al. 2021

Front. Cell Dev. Biol.

View full text Add to dashboard Cite

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.

show abstract

“…Many experimental and computational studies aiming at identifying the channel-opening mechanism of nAChR have been done so far, providing important insight into this process (54). A quaternary twist model was suggested based on normal mode analysis (NMA) calculations with a7 ECD-TMD homology models (17,55).…”

Section: Our Model Of Opening Is Consistent With Previous Experimental and Computational Studiesmentioning

confidence: 99%

A Computational Analysis of the Factors Governing the Dynamics of α7 nAChR and Its Homologs

Gulsevin

Meiler

Horenstein

2020

Biophysical Journal

Self Cite

View full text Add to dashboard Cite

The a7 nicotinic acetylcholine receptor is a homopentameric ion channel from the Cys-loop receptor superfamily targeted for psychiatric indications and inflammatory pain. Molecular dynamics studies of the receptor have focused on residue mobility and global conformational changes to address receptor function. However, a comparative analysis of a7 with its homologs that cannot trigger channel opening has not been made so far. To identify the residues involved in a7 activation, we ran triplicate 500-ns molecular dynamics simulations with an a7 extracellular domain homology model and two acetylcholine-binding protein homologs. We tested the effect of ligand binding and amino acid sequence on the structure and dynamics of the three proteins. We found that mobile regions identified based on root mean-square deviation and root mean-square fluctuation values are not always consistent among the individual a7 extracellular domain simulations. Comparison of the replica-average properties of the three proteins based on dynamic cross-correlation maps showed that ligand binding affects the coupling between the C-loop and the Cys-loop, vestibular loop, and b1-b2 loops. In addition, the main-immunogenic-region-like domain of a7 went through correlated motions with multiple domains of the receptor. These correlated motions were absent or diminished in a7 homologs, suggesting a unique role in a7 activation.

show abstract

Nicotinic receptor pharmacology in silico: Insights and challenges

Cited by 8 publications

References 151 publications

SLURP-1 Controls Growth and Migration of Lung Adenocarcinoma Cells, Forming a Complex With α7-nAChR and PDGFR/EGFR Heterodimer

SLURP-1 Controls Growth and Migration of Lung Adenocarcinoma Cells, Forming a Complex With α7-nAChR and PDGFR/EGFR Heterodimer

Human Three-Finger Protein Lypd6 Is a Negative Modulator of the Cholinergic System in the Brain

A Computational Analysis of the Factors Governing the Dynamics of α7 nAChR and Its Homologs

Contact Info

Product

Resources

About