Ionchannels and channelopaties in the heart Viktria Szts Csatorna mkds Tbb betegsg Drugok kapcsolodsa csat.hoz Sejtekbe ioncsat.expresszios mdszerek, bemutatsa Action of membrane transport protein ATP-powered pump Ion chanels Transporters ions/s ions/s ions/s Nerbonne et al. Circ Res. 2001;89: Membrane topology of the Kv and Kir2.x K-ionchannels H5 Voltage gated K + channel Inward rectifier K + channel Kv channel CO 2 Kv complex N N C C KChAP PSD MiRP Abott et al Neuropharm Klnbz Ioncsatorna alegysgek sszeszereldse Voltage-Gated Calcium Channels Voltage-gated calcium channels are heteromultimers composed of an 1 subunit and three auxiliary subunits, 2-, and . The 1 subunit forms the ion pore and possesses gating functions and, in some cases, drug binding sites. Ten 1 subunits have been identified, which, in turn, are associated with the activities of the six classes of calcium channels. L-type channels have 1C (cardiac), 1D (neuronal/endocrine), 1S (skeletal muscle), and 1F (retinal) subunits; The 1 subunits each have four homologous domains (I-IV) that are composed of six transmembrane helices. The fourth transmembrane helix of each domain contains the voltage-sensing function. The four 1domains cluster in the membrane to form the ion pore. The -subunit is localized intracellularly and is involved in the membrane trafficking of 1subunits. The -subunit is a glycoprotein having four transmembrane segments. The 2 subunit is a highly glycosylated extracellular protein that is attached to the membrane-spanning d-subunit by means of disulfide bonds. The 2-domain provides structural support required for channel stimulation, while the domain modulates the voltage-dependent activation and steady-state inactivation of the channel. Molecular assembly of ion channels CavKvKir Activation and Inactivation Of The Sodium Channel Sodium channels are characterized by voltage- dependent activation, rapid inactivation, and selective ion conductance. Depolarization of the cell membrane opens the ion pore allowing sodium to passively enter the cell down its concentration gradient. The increase in sodium conductance further depolarizes the membrane to near the sodium equilibrium potential. Inactivation of the sodium channel occurs within milliseconds, initiating a brief refractory period during which the membrane is not excitable. The mechanism of inactivation has been modeled as a "hinged lid" or "ball and chain", where the intracellular loop connecting domains III and IV of the a subunit closes the pore and prevents passage of sodium ions. Abriel H. et al., Swiss Med Wkly 2004, w. chwww.sm Ionic currents and ion transporters responsible for cardiac action potential Proposed cellular mechanism for the development of Torsade de pointes in the long QT syndrome Risk factors for developing Torsade de pointes Abriel H. et al., Swiss Med Wkly 2004, Genetic variants (polymorphysm or mutations) Ionic current, proteins and genes associated with inherited arrhythmias Napolitano et al. Pharm. & ther. 2006,110:1-13 Congenital and aquired forms of long QT syndromes Abriel H. et al., Swiss Med Wkly 2004, w. chwww.sm K+, Na+ channel LQT-associated genes and proteins LQT3 Brugada Syndrome, Cardiac conduction defect, Sick sinus syndrome SCN5AI Na LQT7 Andersen-Tawil Syndrome Kir2.1 (KCNJ2)I k1 LQT8 Timothy Syndrome Cav1.2 (CACNA1c)I CaL Kir6.2I kATP Kir3.4I kAch Progressziv familial heart Block1 Kv1.7(KCNA7),Kv1.5I kur LQT2 LQT6, FAF HERG (KCNH2) MiRP1 (KCNE2) I Kr LQT1, JLN1 LQT5, JLN2 KvLQT1(KCNQ1) Mink (KCNE1) I Ks LQTKv4.3I To1 DiseaseGenesCurrent Prolonged QT interval on ECG (reflects prolonged APD) APD governed by a delicate balance between inward (Na + or Ca + ) and outward (K + ) ionic current Affecting the Na + or Ca + channel prolong APD viagain-off- functionmechanism, while mutation in genes encoding K + channel by loss-off-function mechanism Gene mutations in LQT1 and LQT2 LQT1 LQT2 HERG KCNH2 KvLQT1 KCNQ1 Molecular structure and the membrane topology of the HERG channel Mutations in HERG channel Pivotal role of Ser phosphorilation as a regulatory mechanism in Cav1.2 mode1/mode2 gating. Timothys syndrome ShortQT HERG (KCNH2) Kir2.x (KCNJ2) KvLQT1(KCNQ1) I Kr I K1 I Ks Kv3.1, Kv3.3 DiseaseGenesCurrent I Ca CASQ2 (Calsequestrin2) CPVT CPVT catecholamine-induced polymorphic ventricular tachycardia RyR2 CPVT 1 -adrenoceptor ( 1 -AR) 2 -adrenoceptor ( 2 -AR) Risk factor, modify disease or influence progression of disease Risk factor, modify disease or influence progression of disease AF Complexity of protein-protein interaction in cardiomyocytes Missense mutation in calsequestrin2 (CASQ2) Associated with autosomal recessive catecholamine- induced polymorphic ventricular tachycardia (CPVT) Syncope Seizures or Sudden death In response to Physical activity or Emotional stress wild type mutant Kir2.1 ionchannel has an autosomal dominant mutation in Andersen-Tawil Syndrome Cardiac arrhytmias Periodic paralysis Dysmorphic bone structure(scoliosis, low-set ears, small chin, broad forehead Facial and sceletal features in Andersen- Tawil syndrome GIRK mutation ANP role Gene-specific mutation study Genexpression study Microarray, qRT-PCR Proteomica Kir2.x analysis by RT-PCR RV LV RA LA DOG HUMAN n=12n= 6 kDa Expression of Kv1.5 protein in human and dog Co-localization of Kv 2 auxillary subunit with Kv1.5 in dog left ventricular myocytes 100 Kv1.5-FITC Kv 2-Texas red Kv1.5-FITC Kv 2-Texas red