Binding site of AChR: Difference between revisions
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The X-ray structure of AChR has not yet been solved since its hydrophobic character hampers its successful crystallization. So in this page,<ref>PMID:11683996</ref> We will use a complex of α-bungarotoxinwith a high affinity 13-residue peptide that is homologous to the αsubunit of AChR to study the AChR binding site in general. We also will present the [http://proteopedia.org/wiki/index.php/Acetylcholine_binding_protein Acetylcholine binding protein] and the general [http://proteopedia.org/wiki/index.php/4hfi pentameric ligand gated ion channels] to help you understand this kind of structure and their function. | The X-ray structure of AChR has not yet been solved since its hydrophobic character hampers its successful crystallization. So in this page,<ref>PMID:11683996</ref> We will use a complex of α-bungarotoxinwith a high affinity 13-residue peptide that is homologous to the αsubunit of AChR to study the AChR binding site in general. We also will present the [http://proteopedia.org/wiki/index.php/Acetylcholine_binding_protein Acetylcholine binding protein] and the general [http://proteopedia.org/wiki/index.php/4hfi pentameric ligand gated ion channels] to help you understand this kind of structure and their function. | ||
The nAChR is unable to bind ACh when bound to any of the snake venom α-neurotoxins. These α-neurotoxins antagonistically bind tightly and noncovalently to nAChRs of skeletal muscles, thereby blocking the action of ACh at the postsynaptic membrane, inhibiting ion flow and leading to paralysis and death. The nAChR contains two binding sites for snake venom neurotoxins. | Nicotinic receptors, with a molecular mass of 290 kDa,<ref>PMID:15701510</ref> are made up of five subunits, arranged symmetrically around a central pore.Each subunit comprises four transmembrane domains with both the N- and C-terminus located extracellularly. | ||
As with all ligand-gated ion channels, opening of the nAChR channel pore requires the binding of a chemical messenger. Several different terms are used to refer to the molecules that bind receptors, such as ligand. As well as the endogenous agonist acetylcholine, agonists of the nAChR are nicotine, epibatidine, and choline. | |||
In muscle-type nAChRs, the acetylcholine binding sites are located at the α and either ε or δ subunits interface (or between two α subunits in the case of homomeric receptors) in the extracellular domain near the N terminus.When an agonist binds to the site, all present subunits undergo a conformational change and the channel is opened<ref>PMID:15165738</ref> and a pore with a diameter of about 0.65 nm opens. | |||
Nicotinic AChRs may exist in different interconvertible conformational states. Binding of an agonist stabilises the open and desensitised states. Opening of the channel allows positively charged ions to move across it; in particular, sodium enters the cell and potassium exits. The net flow of positively-charged ions is inward. | |||
The nAChR is a non-selective cation channel, meaning that several different positively charged ions can cross through.It is permeable to Na+ and K+, with some subunit combinations that are also permeable to Ca2+<ref>PMID:12761283</ref><ref>PMID:15644873</ref> The amount of sodium and potassium the channels allow through their pores (their conductance) varies from 50–110 pS, with the conductance depending on the specific subunit composition as well as the permeant ion.<ref>PMID:2423878</ref> | |||
The nAChR is unable to bind ACh when bound to any of the snake venom α-neurotoxins. These α-neurotoxins antagonistically bind tightly and noncovalently to nAChRs of skeletal muscles, thereby blocking the action of ACh at the postsynaptic membrane, inhibiting ion flow and leading to paralysis and death. The nAChR contains two binding sites for snake venom neurotoxins. Some studies have shown that a twist-like motion caused by ACh binding is likely responsible for pore opening, and that one or two molecules of α-bungarotoxin (or other long-chain α-neurotoxin) suffice to halt this motion. The toxins seem to lock together neighboring receptor subunits, inhibiting the twist and therefore, the opening motion.<ref>PMID:18327915</ref> | |||
The activation of receptors by nicotine modifies the state of neurons through two main mechanisms. On one hand, the movement of cations causes a depolarizationof the plasma membrane (which results in an excitatory postsynaptic potential in neurons), but also by the activation of voltage-gated ion channels. On the other hand, the entry of calcium acts, either directly or indirectly, on different intracellular cascades leading, for example, to the regulation of the activity of some genes or the release of neurotransmitters. | |||
Revision as of 02:13, 23 January 2015
IntroductionThere are two kinds of acetylcholine receptor in nature: nicotinic acetylcholine receptors and muscarinic acetylcholine receptors. The nicotinic acetylcholine receptor(nAChR) is a pentameric ligand-gated ion channel activated by binding of acetylcholine in nature. In this page we will show the binding site of nAChR. Pentameric ligand-gated ion channelPentameric ligand gated ion channels(), or Cys-loop receptors,are a group of transmembrane ion channel proteins which open to allow ions such as Na+, K+, Ca2+, or Cl- to pass through the membrane in response to the binding of a chemical messenger, such as a neurotransmitter[1]. Nicotinic acetylcholine receptor is a kind of pentameric ligand gated ion channels. So at first of this page, we introduce some facts of the pentameric ligand gated ion channels, which will help us to understand the structure and function of AChR. In overall organization, the pLGIC have five subunits. The five subunits are arranged in a barrel-like manner around a central symmetry axis that coincides with the ion permeation pathway.[2] In each subunit, the extracellular domin(ECD) of pLGIC encompasses 10β-strands that are organized as a sandwich of two tightly interacting β-sheets, while the transmembrane domain(TMD) folds into a bundle of four α-helices. Structure of Acetylcholine binding siteThe X-ray structure of AChR has not yet been solved since its hydrophobic character hampers its successful crystallization.[3] But the X-ray structure of an acetylcholine binding protein() has been solved(Brejc et al., 2001). AChBP is most closely related to the α-subunits of the nAChR. Nearly all residues that are conserved within the nAChR family are present in AChBP, including those that are relevant for lignad binding.[4] We already know that the ligand binding site of AChR is mainly located at the α-subunits. And AChBP can bind with α-Neurotoxins, so AChBP can be use as an example to study the structure of AChR. The high affinity and specific interaction of α-bungarotoxin () with AChR has been of considerable importance in the study of the binding site of AChR.[5] There is a 13-mer high affinity peptides() which corresponding to residues 187-199 of the AChR that can inhibits the binding of α-BTX eo AChR. And through the crystal structure we can study the structure binding site of AChR. The 13-mer HAP assumes an antiparallel β hairpin structure, and is held snugly between of α-BTX. The shortest and most numerous interactions are formed with finger 2 of α-BTX. The intermolecular interaction between finger2 and two arms of the HAP hairpin make the complex stable, like . Affinity labeling experiments which indentified position 10 and 33 of -neurotoxin to be with in 11.5-15.5 Å from AChR residues Cys192-Cys193(Michalet et al ., 2000) agree with the α-BTX-HAP structure where the corresponding Cα distance are 11.05 Å(Pro10-Ser193), 12.72 Å(Cys33-Ser193), 14.45 Å（Pro10-Ser 192）,9.76 Å(Cys33-Ser192),respectively.[6]So through the complex of α-BTX-HAP, we can see the structure of Acetylcholine binding site. Superimpose HAP on AChBPThe crystal shows it's a pentamer like the AChR molecule,which is obviously an ideal candidate for testing the relevance of the conformation of the HAP(The small brown loop binding to blue subunit is the HAP.) when bound to α-BTX, to that of the corresponding binding region in AChR.And the Ach binding site in AChBP is assigned to the 187-199 loop of the AChR αsubunit. α-BTX binds perpendicular to the 5-fold axis of the AChBP molecule and therefore, there are no steric hindrance limitations even when five toxin molecules bind to AChBP.The X-ray confirm that the major interaction between α-BTX and the HAP occur residues 187-192 of AchR α subunit.The overlay of the first 12 residues of the short 13-mer binding HAP assumes a structure similar to the corresponding region of AChR upon binding to α-BTX. The superimposed model of AchBP and α-BTX shows residues 34–36 (corresponding to residues 36–38 of AChR σ subunit) and 162–165 (181–184) of the neighboring AChBP subunit (subunit B) as abutting the α-BTX molecule. File:Combined model of α-BTX-HAP and AchBP.png This figure(Michal Harel,Joel Sussman,2001) shows the stereo view of the combined model of α-BTX-HAP(Red) and AChBP structure with subunit A in green and subunit B in yellow showing the insertion of loop 2 of the toxin into the interface of the to subunits. Function of Acetylcholine receptorThe α-Neurotoxins such as α-bungarotoxin (α-BTX)can compete antagonists of acetylcholine for its site. So studying the binding site of AChR is very important for the development of antidotesagainstα-BTX poisoning as well as drugs against, like Alzheimer's disease and nicotine addiction. The X-ray structure of AChR has not yet been solved since its hydrophobic character hampers its successful crystallization. So in this page,[7] We will use a complex of α-bungarotoxinwith a high affinity 13-residue peptide that is homologous to the αsubunit of AChR to study the AChR binding site in general. We also will present the Acetylcholine binding protein and the general pentameric ligand gated ion channels to help you understand this kind of structure and their function. Nicotinic receptors, with a molecular mass of 290 kDa,[8] are made up of five subunits, arranged symmetrically around a central pore.Each subunit comprises four transmembrane domains with both the N- and C-terminus located extracellularly. As with all ligand-gated ion channels, opening of the nAChR channel pore requires the binding of a chemical messenger. Several different terms are used to refer to the molecules that bind receptors, such as ligand. As well as the endogenous agonist acetylcholine, agonists of the nAChR are nicotine, epibatidine, and choline. In muscle-type nAChRs, the acetylcholine binding sites are located at the α and either ε or δ subunits interface (or between two α subunits in the case of homomeric receptors) in the extracellular domain near the N terminus.When an agonist binds to the site, all present subunits undergo a conformational change and the channel is opened[9] and a pore with a diameter of about 0.65 nm opens. Nicotinic AChRs may exist in different interconvertible conformational states. Binding of an agonist stabilises the open and desensitised states. Opening of the channel allows positively charged ions to move across it; in particular, sodium enters the cell and potassium exits. The net flow of positively-charged ions is inward. The nAChR is a non-selective cation channel, meaning that several different positively charged ions can cross through.It is permeable to Na+ and K+, with some subunit combinations that are also permeable to Ca2+[10][11] The amount of sodium and potassium the channels allow through their pores (their conductance) varies from 50–110 pS, with the conductance depending on the specific subunit composition as well as the permeant ion.[12] The nAChR is unable to bind ACh when bound to any of the snake venom α-neurotoxins. These α-neurotoxins antagonistically bind tightly and noncovalently to nAChRs of skeletal muscles, thereby blocking the action of ACh at the postsynaptic membrane, inhibiting ion flow and leading to paralysis and death. The nAChR contains two binding sites for snake venom neurotoxins. Some studies have shown that a twist-like motion caused by ACh binding is likely responsible for pore opening, and that one or two molecules of α-bungarotoxin (or other long-chain α-neurotoxin) suffice to halt this motion. The toxins seem to lock together neighboring receptor subunits, inhibiting the twist and therefore, the opening motion.[13] The activation of receptors by nicotine modifies the state of neurons through two main mechanisms. On one hand, the movement of cations causes a depolarizationof the plasma membrane (which results in an excitatory postsynaptic potential in neurons), but also by the activation of voltage-gated ion channels. On the other hand, the entry of calcium acts, either directly or indirectly, on different intracellular cascades leading, for example, to the regulation of the activity of some genes or the release of neurotransmitters.
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ReferencesReferences
- ↑ Purves, Dale, George J. Augustine, David Fitzpatrick, William C. Hall, Anthony-Samuel LaMantia, James O. McNamara, and Leonard E. White (2008). Neuroscience. 4th ed. Sinauer Associates. pp. 156–7. ISBN 978-0-87893-697-7.
- ↑ Gonzalez-Gutierrez G, Cuello LG, Nair SK, Grosman C. Gating of the proton-gated ion channel from Gloeobacter violaceus at pH 4 as revealed by X-ray crystallography. Proc Natl Acad Sci U S A. 2013 Oct 28. PMID:24167270 doi:http://dx.doi.org/10.1073/pnas.1313156110
- ↑ Harel M, Kasher R, Nicolas A, Guss JM, Balass M, Fridkin M, Smit AB, Brejc K, Sixma TK, Katchalski-Katzir E, Sussman JL, Fuchs S. The binding site of acetylcholine receptor as visualized in the X-Ray structure of a complex between alpha-bungarotoxin and a mimotope peptide. Neuron. 2001 Oct 25;32(2):265-75. PMID:11683996
- ↑ Brejc K, van Dijk WJ, Klaassen RV, Schuurmans M, van Der Oost J, Smit AB, Sixma TK. Crystal structure of an ACh-binding protein reveals the ligand-binding domain of nicotinic receptors. Nature. 2001 May 17;411(6835):269-76. PMID:11357122 doi:10.1038/35077011
- ↑ Harel M, Kasher R, Nicolas A, Guss JM, Balass M, Fridkin M, Smit AB, Brejc K, Sixma TK, Katchalski-Katzir E, Sussman JL, Fuchs S. The binding site of acetylcholine receptor as visualized in the X-Ray structure of a complex between alpha-bungarotoxin and a mimotope peptide. Neuron. 2001 Oct 25;32(2):265-75. PMID:11683996
- ↑ Harel M, Kasher R, Nicolas A, Guss JM, Balass M, Fridkin M, Smit AB, Brejc K, Sixma TK, Katchalski-Katzir E, Sussman JL, Fuchs S. The binding site of acetylcholine receptor as visualized in the X-Ray structure of a complex between alpha-bungarotoxin and a mimotope peptide. Neuron. 2001 Oct 25;32(2):265-75. PMID:11683996
- ↑ Harel M, Kasher R, Nicolas A, Guss JM, Balass M, Fridkin M, Smit AB, Brejc K, Sixma TK, Katchalski-Katzir E, Sussman JL, Fuchs S. The binding site of acetylcholine receptor as visualized in the X-Ray structure of a complex between alpha-bungarotoxin and a mimotope peptide. Neuron. 2001 Oct 25;32(2):265-75. PMID:11683996
- ↑ Unwin N. Refined structure of the nicotinic acetylcholine receptor at 4A resolution. J Mol Biol. 2005 Mar 4;346(4):967-89. Epub 2005 Jan 25. PMID:15701510 doi:10.1016/j.jmb.2004.12.031
- ↑ Colquhoun D, Sivilotti LG. Function and structure in glycine receptors and some of their relatives. Trends Neurosci. 2004 Jun;27(6):337-44. PMID:15165738 doi:http://dx.doi.org/10.1016/j.tins.2004.04.010
- ↑ Beker F, Weber M, Fink RH, Adams DJ. Muscarinic and nicotinic ACh receptor activation differentially mobilize Ca2+ in rat intracardiac ganglion neurons. J Neurophysiol. 2003 Sep;90(3):1956-64. Epub 2003 May 21. PMID:12761283 doi:http://dx.doi.org/10.1152/jn.01079.2002
- ↑ Weber M, Motin L, Gaul S, Beker F, Fink RH, Adams DJ. Intravenous anaesthetics inhibit nicotinic acetylcholine receptor-mediated currents and Ca2+ transients in rat intracardiac ganglion neurons. Br J Pharmacol. 2005 Jan;144(1):98-107. PMID:15644873 doi:http://dx.doi.org/10.1038/sj.bjp.0705942
- ↑ Mishina M, Takai T, Imoto K, Noda M, Takahashi T, Numa S, Methfessel C, Sakmann B. Molecular distinction between fetal and adult forms of muscle acetylcholine receptor. Nature. 1986 May 22-28;321(6068):406-11. PMID:2423878 doi:http://dx.doi.org/10.1038/321406a0
- ↑ Samson AO, Levitt M. Inhibition mechanism of the acetylcholine receptor by alpha-neurotoxins as revealed by normal-mode dynamics. Biochemistry. 2008 Apr 1;47(13):4065-70. doi: 10.1021/bi702272j. Epub 2008 Mar 8. PMID:18327915 doi:http://dx.doi.org/10.1021/bi702272j