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by: Rico Leuschke
Rico Leuschke

GPA 3.7

R. Vogt

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R. Vogt
Class Notes
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This 2 page Class Notes was uploaded by Rico Leuschke on Monday October 26, 2015. The Class Notes belongs to BIOL 635 at University of South Carolina - Columbia taught by R. Vogt in Fall. Since its upload, it has received 32 views. For similar materials see /class/229625/biol-635-university-of-south-carolina-columbia in Biological Sciences at University of South Carolina - Columbia.

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Date Created: 10/26/15
Outline Neurobiology 635 Action Potential 2 Molecular Biology of Voltage Sensitive Ion Channels I Review NernstGoldman Changing Permeabilities Voltage sensitive ion channels II Na Channel isolation cloning and structure A Source electric fish electroplax organ BCloned Na channel Noda et al 1984 purify protein from elec fish TTX binding assay partially sequence protein make antibody to purified protein screen expression cDNA library using antibody sequence cDNA con rm sequence against partial protein sequence analyze for transmembrane domains structural model U ePJN 0quot HI General Schem of Voltage Sensitive Ion Channels A 4 Subunits 1 Na gene encodes single protein containing all 4 subunits domains 2 K genes encode smaller protein corresponding to one domain channel is a tetramer of 4 peptides analagous to hemoglobin but in membrane 3 Ca voltage sensitive ion channel like Na channel single gene encodes all 4 subunits B Features 1 transmembrane regions 2 pore region ion selectivity 3 S4 gating mechanism voltage sensitivity 4 N terminus self inactivation C Multiple Conformational States Descrete stable conformations shapes Unstable intermediates analogy light switch stable on or off unstable inbetween IV K Channel Cloning A Drosophila Genetics B Shaker Mutant Kaplan amp Trout 1969 Trout amp Kaplan 1973 C Solkoffamp Wyman 1981ab 0 Review of Voltage Clamp 1 Development of quotAquot and quotDRquot currents 2 Identification that Shaker Mutant is quotAquot current mutants D Timpe et al 1988 Cloning and expression of K channel in Xenopus frog oocytes V Next Analysis of cloned ion channels Molecular Biology of Ion Chanels I Noda M S Shimizu T Tanabe T Takai T Kayano T lkeda H Takahashi H Nakayama Y Kanaoka N Minamino K Kangawa H Matsuo MA Raftery T Hirose S Inayama H Hayashida T Miyata amp S Numa 1984 Primary structure of Electrophorus electricus sodium channel deduced from cDNA sequence Nature 312 121127 Kaplan WD amp WE Trout 1969 The behavior of four neurological mutants of Drosophila Genetics 61 399409 Shaker diethylether Trout WE amp WD Kaplan 1973 Genetic manipulations of motor output in Shaker mutants of Drosophila J Neurobiol 4 495512 Slakoff L amp R Wyman 1981 Outward currents in developing Drosophila ight muscle Science 212 461463 Salkoff L amp R Wyman 1981 Genetic modification of potassium channels in Drosophila Shaker mutants Nature 293 228230 Salkoff LB amp RJ Wyman 1983 Ion currents in Drosophila ight muscles J Physiol 337 687709 Schwarz TL BL Tempel DM PapaZian YN Jan amp LY Jan 1988 Multiple potassiumchannel components are produced by alternative splicing at the Shaker locus in Drosophila Nature 331 137142 sequenced multiple cDNA clones and compared sequences with genomic sequence Timpe LC TL Schwalz BL Tempel DM PapaZian Y Nung amp LY Jan 1988 Expression of functional potassium channels from Shaker cDNA in Xenopus oocytes Nature 331 143145 express and compare currents of two expressed alt spliced clones Pongs O 1992 Structural basis of voltagegated K channel pharmacology Trends in Pharmacological Sciences 13 359365 Jan LY amp Jan YN 1997 Cloned potassium channels from eukaryotes and prokaryotes Annu Rev Neurosci 20 91123 includes K leakage channels Grigoriev NG Spafford JD Gallin WJ Spencer AN 1997 Voltage sensing in jellyfish Shaker K channels J Exp Biology 200 29192926 Jentsch TJ amp Gunther W 1997 Chloride channels an emerging molecular picture BioEssays 19 1 17126


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