Substrate and Product Trafficking through the Active Center Gorge of Acetylcholinesterase Analyzed by Crystallography and Equilibrium Binding

Bourne, Yves; Radić, Zoran; Sulzenbacher, G.; Kim, Esther; Taylor, Palmer; Marchot, Pierre

doi:10.1074/jbc.m603018200

Cited by 123 publications

(145 citation statements)

References 62 publications

Supporting

Mentioning

139

Contrasting

Order By: Relevance

“…The residues of Trp86, Gly122, Ser203, Phe295, Phe297, and His447 are actively participating in VdW contacts, which are involved in substrate trafficking of ACh to the catalytic site. All these interactions and distance values are in good agreement with that of reported crystal structure of ACh/mAChE (mouse AChE) complex [17]. However, in Lig_6-bound hAChE-ACh complex, the substrate molecule is positioned in PAS site instead of binding with CT region of hAChE due to the active participation of Lig_6 in both PAS and CAS site with the GOLD score of 34.163.…”

Section: Docking Studies Of Hache With Its Agonists (Ach and Aβ)supporting

confidence: 88%

“…The N3 (P pKa 07.78) of this ligand ideally positioned for a hydrogen bonding with OG group of Ser125 with distance of 3.08 Å. The nitro group establishes three HBs, in which O5 forms two HBs with OH of Tyr124 (P p K a 018.40) (2.88 Å), and Tyr337 (P p K a 018.25) (2.97 Å), which is a mediator residue for the mid-point of the diameter gorge [17] and responsible for the regulation of substrate and product trafficking. Atom O6 of the nitro group forms a hydrogen bond with Tyr124 with a distance of 2.75 Å.…”

Section: Docking Of Lig_1 To Pas Of Hachementioning

confidence: 99%

“…The methylsulfanyl group faces the six-membered ring of the Trp86 indole by the active C-H-π interaction with a distance of 3.23 Å, which is similar to cation-π interaction as observed in the ACh-hAChE complex. The benzyl group of this ligand forms two adjacent site-stacked interaction with Tyr124 by the distance of 4.09 Å (Tyr124 CZ-C12 Lig_1) and 3.87 Å (Tyr124 CE1-C13 Lig_1), which is crucial residues of PAS site and helping for substrate binding and trafficking [17]. These results reveal that this ligand can span the entire gorge of hAChE, which might attribute to the inhibition of Aβ deposition and to the reduction in the hydrolysis of ACh (Fig.…”

Section: Docking Of Lig_1 To Pas Of Hachementioning

confidence: 99%

“…Most of the administered drugs interact with the CT of AChE [15,16]. Furthermore, their interaction with residues (Trp86 and Tyr337) of catalytic anionic subsite (CAS), placed in close proximity to the active site, influences the hydrolysis of ACh [9,13,[17][18][19]. Besides the CT and CAS, the residues of peripheral anionic site (PAS; Tyr72, Asp74, Tyr124, Trp286, and Tyr341) also influence the function of AChE through their allosteric effects [16,20,21].…”

Section: Introductionmentioning

confidence: 99%

“…Besides the CT and CAS, the residues of peripheral anionic site (PAS; Tyr72, Asp74, Tyr124, Trp286, and Tyr341) also influence the function of AChE through their allosteric effects [16,20,21]. Residues of PAS trap the substrate molecules as they enter the gorge, and are involved in accelerating the transfer of the substrate acyl group to a serine hydroxyl group in CAS [17,18,[22][23][24][25]. In addition, PAS site is potentially responsible for colocalization of amyloid-β (Aβ) peptide in the brains of AD patients and has been shown to accelerate the assembly of Aβ to form amyloid fibrils [26][27][28].…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

Synthesis and in silico evaluation of 1N-methyl-1S-methyl-2-nitroethylene (NMSM) derivatives against Alzheimer disease: to understand their interacting mechanism with acetylcholinesterase

et al. 2012

View full text Add to dashboard Cite

Anomalous action of human acetylcholinesterase (hAChE) in Alzheimer's disease (AD) was restrained by various AChE inhibitors, of which the specific and potent lead candidate Donepezil is used for treating the disease AD. Besides the specificity, the observed undesirable side effects caused by Donepezil invoked the quest for new lead molecules with the increased potency and specificity for AChE. The present study elucidates the potency of six 1N-methyl-1S-methyl-2-nitroethylene (NMSM) derivatives to form a specific interaction with the peripheral anionic site and catalytic anionic subsite residues of hAChE. The NMSMs were prepared in good yield from 1,1-di(methylsulfanyl)-2-nitroethylene and primary amine (or) amino acid esters. In silico interaction analysis reveals specific and potent interactions between hAChE and selected ligand molecules. The sitespecific interactions formed between these molecules also results in a conformational change in the orientation of active site residues of hAChE, which prevents them from being accessed by beta-amyloid protein (Aβ), which is a causative agent for amyloid plaque formation and acetylcholine (ACh). In silico interaction analysis between the ligand-bounded hAChE with Aß and ACh confirms this observation. The variation in the conformation of hAChE associated with the decreased ability of Aβ and ACh to access the respective functional residues of hAChE induced by the novel NMSMs favors their selection for in vivo analysis to present themselves as new members of hAChE inhibitors.

show abstract

Section: Docking Studies Of Hache With Its Agonists (Ach and Aβ)supporting

confidence: 88%

Section: Docking Of Lig_1 To Pas Of Hachementioning

confidence: 99%

Section: Docking Of Lig_1 To Pas Of Hachementioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Synthesis and in silico evaluation of 1N-methyl-1S-methyl-2-nitroethylene (NMSM) derivatives against Alzheimer disease: to understand their interacting mechanism with acetylcholinesterase

et al. 2012

View full text Add to dashboard Cite

show abstract

Back‐Scattering Interferometry: An Ultrasensitive Method for the Unperturbed Detection of Acetylcholinesterase–Inhibitor Interactions

et al. 2012

View full text Add to dashboard Cite

A series of inhibitors of acetylcholinesterase (AChE) have been screened by back-scattering interferometry (BSI). Enzyme levels as low as 100 pM (22,000 molecules of AChE) can be detected. This method can be used to screen for mixed AChE inhibitors, agents that have shown high efficacy against Alzheimer's disease, by detecting dual-binding interactions. Keywords detection method; interferometry; acetylcholinesterase; high-throughput screening; enzyme assayIn recent years, the emphasis of drug discovery and optimization has shifted from traditional methods to rapid microfluidic screening to minimize cost and maximize output. Backscattering interferometry (BSI),[1] a technique which quantifies refractive index (RI) changes arising from intermolecular binding interactions, is a novel biosensing platform that may be useful in drug discovery. Acetylcholinesterase (AChE), a widely studied serine hydrolase that plays pivotal roles in Alzheimer's disease (AD), [2] inflammatory processes, [3] and nerve-agent poisoning, [4] is an interesting system with which to study the use of BSI for the rapid detection of drug candidates. The search for potent AChE inhibitors (AChEI) has been driven largely by the need for an effective AD treatment, and is based on the longstanding cholinergic hypothesis [2] and the more recent amyloid hypothesis. [5] It is wellestablished that AChE accelerates amyloid-beta (Ab) peptide deposition, a process which may be mediated by an interaction between plaque precursors and the peripheral anionic site (PAS) of the enzyme.[6] Recent drug discovery efforts have focused on the design of AChEIs that are able to interact with the PAS to target both cholinergic and noncholinergic AD pathologies. [7] Seminal work in this field has led to the discovery of dual-binding inhibitors, such as bisgalantamine [8] and pseudo-irreversible carbamate inhibitors. [9] Given the large number of studies on several diverse classes of AChEIs, this system is a widely accepted benchmark for the development and testing of novel screening techniques.Traditional methods used to detect interactions with AChE focus on the indirect measurement of substrate procedures. [10] To explore the utility of BSI to detect AChEIs, several known and novel inhibitors with diverse potencies and inhibitory mechanisms have been screened. Herein, we show that BSI can: 1) quantify AChE-inhibitor interactions in the We first wanted to probe the utility of BSI to screen AChEIs using a set of known and novel AChEIs (Scheme 1). Edrophonium, a competitive inhibitor, propidium, a noncompetitive inhibitor, and the mixed inhibitors, 1,5-bis(4-allyldimethylammoniumphenyl) pentan-3-one dibromide (BW284c51) and galantamine, were the standards used to validate BSI as a method for the detection of AChEIs. Two separate and distinct saturation curves were observed for inhibitors 4 and 6 (Figure 2 and Supporting Information, Figure S7, respectively). For ligand 4, the dual-binding curve is shown in the graph inset of while the fit of the second bindin...

show abstract

Back‐Scattering Interferometry: An Ultrasensitive Method for the Unperturbed Detection of Acetylcholinesterase–Inhibitor Interactions

et al. 2012

View full text Add to dashboard Cite

show abstract

Substrate and Product Trafficking through the Active Center Gorge of Acetylcholinesterase Analyzed by Crystallography and Equilibrium Binding

Cited by 123 publications

References 62 publications

Synthesis and in silico evaluation of 1N-methyl-1S-methyl-2-nitroethylene (NMSM) derivatives against Alzheimer disease: to understand their interacting mechanism with acetylcholinesterase

Synthesis and in silico evaluation of 1N-methyl-1S-methyl-2-nitroethylene (NMSM) derivatives against Alzheimer disease: to understand their interacting mechanism with acetylcholinesterase

Back‐Scattering Interferometry: An Ultrasensitive Method for the Unperturbed Detection of Acetylcholinesterase–Inhibitor Interactions

Back‐Scattering Interferometry: An Ultrasensitive Method for the Unperturbed Detection of Acetylcholinesterase–Inhibitor Interactions

Contact Info

Product

Resources

About