Ion Channels Presented by- Aindrila Saha Roll Number-1411009 Third Year School of Biological Sciences B351: Principles of Drug DesigningWhat are ion channels? * Pore forming transmembrane proteins associated with transport of specific ions in or out of the cell. * Highly selective in type of ion transported (exceptions are there), * Very high rate of ion transfer. * fons are transported across electrochemical gradient. * Passive mechanism.Discovery * Fundamental properties of channels mediated by currents were first discovered by Alan Hodgkin and Andrew Huxley on their work on action potential (1952). * The existence of ion channels was confirmed in the 1970s by Bernard Katz and Ricardo Miledi using noise analysis. * The Nobel Prize in Chemistry for 2003 was awarded to two American scientists: Roderick MacKinnon for his studies on the physico-chemical properties of ion channel structure and function and andBiological Roles * Conductance of Nerve impulse, generation of action potential, synaptic transmission. * Cardiac, skeletal and smooth muscle contraction. * Epithelial transport of nutrients and ions. * T-cell activation (immune regulation). * Pancreatic beta cell insulin release.Classification * On the basis of gating. - On the basis of type of ion passed. » Number of gates present. * Localization of proteins in the cell.Classification based on gating - Voltage Gated lon Channels - Open and close in response to membrane potential. * Ligand Gated lon Channels - Open in response to specific ligand molecules binding to the extracellular domain of the receptor protein. - Other Gatings - Indirect signalling, mechano-gated ion channels. light cated channels.a) Voltage Gated ion channels * Voltage sensitive * Conformational change in response to the potential gradient. * Generally ion specific. * Important for excitable cells like neurons. * Role in regulation of depolarization and polarization of neuronal membrane during an action potential. * Distributed along the axon and soma of the neurons.Types of Voltage Gated Channels ‘ Voltage Gated Sodium Channels (9 members, responsible for membrane depolarization in action potential generation) * Voltage Gated Calcium Channels (10 members,play an important role in both linking muscle excitation with contraction as well as neuronal excitation with transmitter release. ) * Voltage Gated Potassium Channels (40 members, role in repolarization of cell membrane ofter action potentialTypes of Voltage Gated Channels ‘ Transient receptor potential channels (TRP channels): 28 types, some of them are voltage gated, named after their role in Drosophila phototransduction. * Hyperpolarization-activated cyclic nucleotide-gated channel ( pacemaking channels in the heart, sensitive to cAMP, cGMP that alter the voltage sensitivity of the channels) ‘ Voltage sensitive proton channels (helps in acid extrusion from cell, phaqgocvtosis. stronaly oH reaulated)Structure * Several subunits with a central pore. * lon specific, but ions with similar charge and size can enter. * Functionality governed by 3 main parts- the voltage sensor the pore and the gate. * Na, K and Ca channels have 4 transmembrane alpha subunits surrounding the pore. * Six subunits: S1-S6. S1-S4: Voltage sensing region, S5-S6: Gate and pore. >» Dani ilatran;s hats eishiiniteFig: Subunits of a Voltage gated ion channel Source: By Efazzari - Own work, CC BY-SA 4.0, https://commons.wikimedia.org/wiindex.php? curid=47402794Mechanism of action (ps nce a Agee cpee a MY ey aMechanism ‘ For potassium channel: When a potential difference is introduced over the membrane, the associated electric field induces a conformational change in the potassium channel. The conformational change distorts the shape of the channel proteins sufficiently such that the cavity, or channel, opens to allow influx or efflux to occur across the membrane.Mechanism * Moltage sensing in Na and Ca channels: Positve charges in the voltage sensing domain, Presence of Arginine and histidine repeats in this segment, conserved domain. * Gate acts as a mechanical obstruction to ion flow. * Channel closes milliseconds after opening.b) Ligand Gated Channels * Group of transmembrane ion channels that allow the passing of several ions upon the binding of specific chemical messenger like neurotransmitters. * Two domains: Transmembrane domain including channel pore, Extracellular domain including ligand binding site. * Function: Conversion of presynaptic chemical signal quickly and effectively into post-synaptic electreical signal. * Three super families: cys-loop ee ag Se en aLCys-Loop Receptors * Characteristic loop formed by a disulfide bond between two cysteine residues in the N terminal extracellular domain. * Provides specificity for (1) acetylcholine (AcCh), (2) serotonin, (3) glycine, (4) glutamate and (5) y-aminobutyric acid (GABA) in vertebrates. * Structural elements are well conserved, with a large extracellular domain (ECD) harboring an alpha-helix and 10 beta- strands. Following the ECD, four transmembrane seaqments (TMSs) areIonotropic Glutamate Receptor * Binds to NT Glutamate. * Consists of a tetramer. * Each sub-unit consists of extracellular amino terminal domain (ATD, which is involved in tetramer assembly), an extracellular ligand binding domain (LBD, which binds glutamate), and a transmembrane domain (TMD, which forms the ion channel). * Each subunit of the tetramer has a binding site for glutamate formed by the two LBD oe Re Be oe BR M@B Le mas Bie tea!ATP Gated channels - Bind to ATP in order to open. * They form trimers with two transmembrane helices per subunit and both the C and N_ termini on the intracellular side.[ ~ Mechanism and Receptors lonotropic and Metabotropic Receptors EXTRACELLULAR Neurotransmitter FLUID Neurotransmitter ~ lon channel aL Cell signaling CYTOSOL ta) Direct neurotransmitter action (b) Indirect neurotransmitter action receptor} (metabotropic receptor) __. Source: hitp:/www.mun.calbiology/desmid/brian/BIOL2060/BIOL2060- i 1313_17,jpg ,Classification on the basis of types of ions passed * Chloride Channels * Potassium Channels (Voltage-gated potassium channels, Calcium-activated potassium channels , Inward-rectifier potassium channels, Two-pore-domain potassium channels) * Sodium Channels (Voltage-gated sodium channels, Epithelial sodium channels) - Calcium Channels - Proton Channels (Voltage Gated Proton Channels)Classification on the basis of Localisation * Plasma Membrane Channels : Example- Voltage-gated potassium channels (Kv), Sodium channels (Nav), Calcium channels (Cav) and Chloride channels (CIC) - Intracellular Channels : Example — Endoplasmic Reticular channels (RyR, SERCA, ORAi) and Mitochondrial channels (mPTP, KATP, BK, IK, CLICS5, Kv7.4 at the inner membrane and VDAC and CLIC4 as outer membrane channels.)Ion Chanel Kinetics * In electrophysiology, “Gating” refers to the opening (activation) and closing (inactivation) of ion channels. * Kinetics is modeled by a two state Markov Model with alpha, the activation constant and beta, the inactivation constant. * Single channel kinetics are mostly recorded by Patch clamp technique.Biological Implication * Action Potential Generation (Voltage Gated Channels) ® depolarisation J \etiage-gied Na* channel Jegeeoses tH 2,9, 20 6 af > Source: http://www.vce.bioninja.com.au/aos-2-detecting-and- respond/coordination—regulation/nervous-system.htmlBiological Implications * Ventricular Myosite Action Potential (Voltage Gated Channels) Relationship Between Cardiac Action Potential and lon Channel CurrentsBiological Implications * Synaptic Transmission (Voltage and Ligand Gated Mechanisms) Neurotransmitter Vesicle Calcium channel Receptors } Symaptic let Sodium”) channel |, Piste Source: http://ibguides.com/images/chemical-synapse.pngDrugs Targetting Ion Channels - lon Channels are varied widely and play a wide range of critical biological functions. 55 different medical conditions have been attributed to ion channel dysfunction. - Owing to these conditions, 13.4% of all drugs are targetted to ion channels (second highest after GPCRs). * Worldwide sale of ion channel drugs (estimate) > $12 billion. © Source: Discov Med. 2010 Mar:9(46):253-60, “Targeting ion channels for drug discovery’, Clare JV.Drugs Targetting Ion Channels * 4. Tetrodotoxin * 2. Ziconitide * 3. Benzodiazepines * 4. Conotoxin * §. Lidocaine * Other drugs are: Verapamil, Diltiazem, Amlodipine, Nimodipine, Nifedipin, Lidocaine etc. * Most of them are used as anaesthetics, to cure epilepsy, treat hypertension and1. Tetrodotoxin * Source: Tetraodontiformes, an order that includes pufferfish, porcupinefish, ocean sunfish, and triggerfish. * Structure: 0 OH oe N Structure of Tetrodotoxin Molecule (Source: Wikipedia) IHTetrodotoxin * Pharmacological Activity : Potent Sodium Channel blocker, Neurotoxin and reduces drug craving and anxiety in abstinent heroin addicts. * Biomolecular Target: A partuicular type of fast acting Sodium Channel. * Mechanism of Action: Binds to the site 1 f a specific Volage gated fast opening sodium channel and temporarily blocks the functioning of the channel. TTX-Na+-s (present in most part of the body) and TTX-Na+-r (present in cardiac tissue).2. Ziconitide * Source: Cone snail Conus magus. * Structure: Ziconotide is a peptide with the amino acid sequence H-Cys-Lys-Gly-Lys-Gly-Ala-Lys-Cys-Ser- Arg-Leu-Met-Tyr-Asp-Cys-Cys-Thr-Gly-Ser-Cys-Arg- Ser-Gly-Lys-Cys-NH2.Ziconitide * Pharmacological activity: | Potentic analgesic with 1000 times the potency of Morphine. * Biomolecular target: Specific CaN channel. ~ Route of administration: Intrathecally * Mechanism of Action: a selective N- type(neuronal type) voltage-gated calcium channel blocker. This action inhibits the release of pro-nociceptive neurochemicals like glutamate, calcitonin gene-related ne ope OL3. Benzodiazepines * Structure: Fusion of a benzene ring with a diazepine ring. ~ Source: By Jil - Own work, CC BY-SA. 4.0,hitps://commons. wikimedia.orgiwiindex.php?curid=49864697Benzodiazepine * Pharmacological activity: Acts as sedative, hypnotic, anxiolytic, anticonvulsant, muscle relaxant, used to treat a variety of indications such as alcohol dependence, seizures, anxiety disorders, panic, agitation, and insomnia. * Mechanism of Action: Activates inhibitore neurotransmitter GABA, acts as a positive allosteric modulator of GABAa receptor by increasing the total conduction of chloride ions across the neuronal cell membrane when GABA is already boundBenzodiazepine - Side effects: Generally considered safe for short term use but have withdrawal effects due to long term use. Prescribing during pregnancy is controversial. Elderly at increased risk of short and long term effects.4, Conotoxin * Source: Derived from different species of cone snail (Genus: Conus). * Structure: Peptide with 30-40 amino acid residues and 1-2 disulphide bonds. Source: By Fvasconcellos (talk contribs) - From PDB entry 4AKG., Public Domain, https://commons. wikimedia,org/w/ index.php?curid=4286936Conotoxin * Biomolecular target: Different varieties of conotoxin affects different ion channels and receptors. a-conotoxinc - inhibits nicotinic acetylcholine receptors at nerves and muscles. 5-conotoxin - inhibits the inactivation of voltage-dependent sodium channels. k-conotoxin- inhibits potassium channels. - u-conotoxin -inhibits voltage-dependent sodium channels in muscles.Conotoxin - Mechanism of action and Pharmacological activity: w-conotoxin has an analgesic effect and w-conotoxin M VII A is 100 to 1000 times that of morphine. Synthetic version used as analgesic.5. Lidocaine * Structure: 2-(diethylamino)-N-(2,6- dimethylphenyl)acetamide Source: By Ben Mills - Own work, Public Domain, https://commons.wikim edia.org/wiindex.php? curid=5750147Lidocaine * Pharmacological Activity: Local anaesthetic with rapid onset of action and immediate response, important class-1b antiarrhythmic drug, topically used for neuropathis pain. * Bioavailalbility: 35% (oral), 5% (topical). * Mechanism of Action: Voltage dependent fast sodium channel blocker, prevents polarization of post synaptic membrane and signal conduction.Ion Channel dysfunction and Diseases * Channelopathies * Cystic fibrosis is caused by mutations in the CFTR gene, which is a_ chloride channel. * Brugada syndrome is another ventricular arrhythmia caused by voltage-gated sodium channel gene mutations. * Shaker gene mutations, ataxia etc.mething eres FED TELE, ing to be THANK YOU!