Pharmacological characterization of small‐conductance Ca2+‐activated K+ channels stably expressed in HEK 293 cells
Three genes encode the small‐conductance Ca2+‐activated K+ channels (SK channels). We have stably expressed hSK1 and rSK2 in HEK 293 cells and addressed the pharmacology of these subtypes using whole‐cell patch clamp recordings. The bee venom peptide apamin blocked hSK1 as well as rSK2 with IC50 val...
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| Veröffentlicht in: | British journal of pharmacology 2000-03, Vol.129 (5), p.991-999 |
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| creator | Strøbæk, Dorte Jørgensen, Tino D Christophersen, Palle Ahring, Philip K Olesen, Søren‐Peter |
| description | Three genes encode the small‐conductance Ca2+‐activated K+ channels (SK channels). We have stably expressed hSK1 and rSK2 in HEK 293 cells and addressed the pharmacology of these subtypes using whole‐cell patch clamp recordings.
The bee venom peptide apamin blocked hSK1 as well as rSK2 with IC50 values of 3.3 nM and 83 pM, respectively.
The pharmacological separation between the subtypes was even more prominent when applying the scorpion peptide blocker scyllatoxin, which blocked hSK1 with an IC50 value of 80 nM and rSK2 at 287 pM.
The potent small molecule blockers showed little differentiation between the channel subtypes. The bis‐quinolinium cyclophane UCL 1684 blocked hSK1 with an IC50 value of 762 pM and rSK2 at 364 pM. The antiseptic compound dequalinium chloride blocked hSK1 and rSK2 with IC50 values of 444 nM and 162 nM, respectively.
The nicotinic acetylcholine receptor antagonist d‐tubocurarine was found to block hSK1 and rSK2 with IC50 values of 27 μM and 17 μM when measured at +80 mV. The inhibition by d‐tubocurarine was voltage‐dependent with increasing affinities at more hyperpolarized potentials.
The GABAA receptor antagonist bicuculline methiodide also blocked hSK1 and rSK2 in a voltage‐dependent manner with IC50 values of 15 and 25 μM when measured at +80 mV.
In conclusion, the pharmacological separation between SK channel subtypes expressed in mammalian cells is too small to support the notion that the apamin‐insensitive afterhyperpolarization of neurones is mediated by hSK1.
British Journal of Pharmacology (2000) 129, 991–999; doi:10.1038/sj.bjp.0703120 |
| doi_str_mv | 10.1038/sj.bjp.0703120 |
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The bee venom peptide apamin blocked hSK1 as well as rSK2 with IC50 values of 3.3 nM and 83 pM, respectively.
The pharmacological separation between the subtypes was even more prominent when applying the scorpion peptide blocker scyllatoxin, which blocked hSK1 with an IC50 value of 80 nM and rSK2 at 287 pM.
The potent small molecule blockers showed little differentiation between the channel subtypes. The bis‐quinolinium cyclophane UCL 1684 blocked hSK1 with an IC50 value of 762 pM and rSK2 at 364 pM. The antiseptic compound dequalinium chloride blocked hSK1 and rSK2 with IC50 values of 444 nM and 162 nM, respectively.
The nicotinic acetylcholine receptor antagonist d‐tubocurarine was found to block hSK1 and rSK2 with IC50 values of 27 μM and 17 μM when measured at +80 mV. The inhibition by d‐tubocurarine was voltage‐dependent with increasing affinities at more hyperpolarized potentials.
The GABAA receptor antagonist bicuculline methiodide also blocked hSK1 and rSK2 in a voltage‐dependent manner with IC50 values of 15 and 25 μM when measured at +80 mV.
In conclusion, the pharmacological separation between SK channel subtypes expressed in mammalian cells is too small to support the notion that the apamin‐insensitive afterhyperpolarization of neurones is mediated by hSK1.
British Journal of Pharmacology (2000) 129, 991–999; doi:10.1038/sj.bjp.0703120</description><identifier>ISSN: 0007-1188</identifier><identifier>EISSN: 1476-5381</identifier><identifier>DOI: 10.1038/sj.bjp.0703120</identifier><identifier>PMID: 10696100</identifier><identifier>CODEN: BJPCBM</identifier><language>eng</language><publisher>Oxford, UK: Blackwell Publishing Ltd</publisher><subject>apamin ; bicuculline methiodide ; Biological and medical sciences ; Calcium‐activated potassium channel ; Cell membranes. Ionic channels. Membrane pores ; Cell structures and functions ; dequalinium chloride ; d‐tubocurarine ; Fundamental and applied biological sciences. Psychology ; HEK 293 ; Molecular and cellular biology ; scyllatoxin ; taicatoxin ; UCL 1684</subject><ispartof>British journal of pharmacology, 2000-03, Vol.129 (5), p.991-999</ispartof><rights>2000 British Pharmacological Society</rights><rights>2000 INIST-CNRS</rights><rights>Copyright Nature Publishing Group Mar 2000</rights><rights>Copyright 2000, Nature Publishing Group 2000 Nature Publishing Group</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC1571906/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC1571906/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,723,776,780,881,1411,1427,27903,27904,45553,45554,46388,46812,53770,53772</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=1289725$$DView record in Pascal Francis$$Hfree_for_read</backlink></links><search><creatorcontrib>Strøbæk, Dorte</creatorcontrib><creatorcontrib>Jørgensen, Tino D</creatorcontrib><creatorcontrib>Christophersen, Palle</creatorcontrib><creatorcontrib>Ahring, Philip K</creatorcontrib><creatorcontrib>Olesen, Søren‐Peter</creatorcontrib><title>Pharmacological characterization of small‐conductance Ca2+‐activated K+ channels stably expressed in HEK 293 cells</title><title>British journal of pharmacology</title><description>Three genes encode the small‐conductance Ca2+‐activated K+ channels (SK channels). We have stably expressed hSK1 and rSK2 in HEK 293 cells and addressed the pharmacology of these subtypes using whole‐cell patch clamp recordings.
The bee venom peptide apamin blocked hSK1 as well as rSK2 with IC50 values of 3.3 nM and 83 pM, respectively.
The pharmacological separation between the subtypes was even more prominent when applying the scorpion peptide blocker scyllatoxin, which blocked hSK1 with an IC50 value of 80 nM and rSK2 at 287 pM.
The potent small molecule blockers showed little differentiation between the channel subtypes. The bis‐quinolinium cyclophane UCL 1684 blocked hSK1 with an IC50 value of 762 pM and rSK2 at 364 pM. The antiseptic compound dequalinium chloride blocked hSK1 and rSK2 with IC50 values of 444 nM and 162 nM, respectively.
The nicotinic acetylcholine receptor antagonist d‐tubocurarine was found to block hSK1 and rSK2 with IC50 values of 27 μM and 17 μM when measured at +80 mV. The inhibition by d‐tubocurarine was voltage‐dependent with increasing affinities at more hyperpolarized potentials.
The GABAA receptor antagonist bicuculline methiodide also blocked hSK1 and rSK2 in a voltage‐dependent manner with IC50 values of 15 and 25 μM when measured at +80 mV.
In conclusion, the pharmacological separation between SK channel subtypes expressed in mammalian cells is too small to support the notion that the apamin‐insensitive afterhyperpolarization of neurones is mediated by hSK1.
British Journal of Pharmacology (2000) 129, 991–999; doi:10.1038/sj.bjp.0703120</description><subject>apamin</subject><subject>bicuculline methiodide</subject><subject>Biological and medical sciences</subject><subject>Calcium‐activated potassium channel</subject><subject>Cell membranes. Ionic channels. Membrane pores</subject><subject>Cell structures and functions</subject><subject>dequalinium chloride</subject><subject>d‐tubocurarine</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>HEK 293</subject><subject>Molecular and cellular biology</subject><subject>scyllatoxin</subject><subject>taicatoxin</subject><subject>UCL 1684</subject><issn>0007-1188</issn><issn>1476-5381</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2000</creationdate><recordtype>article</recordtype><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNpVkcFu1DAQhi0EotvClbOFuFVZPHZiOxckumpZ1Er0AGdr4jitI6-9xNmF5cQj8Iw8Ca66AnGyxv83_4zmJ-QVsCUwod_mcdmN2yVTTABnT8gCaiWrRmh4ShaMMVUBaH1CTnMeGSuiap6TE2CylcDYguxv73HaoE0h3XmLgdpSo53d5H_g7FOkaaB5gyH8_vnLptjv7IzROrpCfl6-Cur3OLueXp8_9MboQqZ5xi4cqPu-nVzORfSRri-vKW8FtS6E_II8GzBk9_L4npEvV5efV-vq5tOHj6v3N9UoaimrmkvFNMceh7ardaM66LAu9YDYgmoHVqPoh3oQveTK9VyDHoCr3oFqbN2JM_Lu0Xe76zauty7OEwaznfwGp4NJ6M3_SvT35i7tDTQKWiaLweujwZS-7lyezZh2Uyw7Gw4KNG-5KNCbI4S53HCYyoV8_jsFuG4VbwomHrFvPrjDP5mZhyhNHk2J0hyjNBe3ayG5FH8AL5iWFg</recordid><startdate>200003</startdate><enddate>200003</enddate><creator>Strøbæk, Dorte</creator><creator>Jørgensen, Tino D</creator><creator>Christophersen, Palle</creator><creator>Ahring, Philip K</creator><creator>Olesen, Søren‐Peter</creator><general>Blackwell Publishing Ltd</general><general>Nature Publishing</general><scope>IQODW</scope><scope>3V.</scope><scope>7QP</scope><scope>7RV</scope><scope>7TK</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M7P</scope><scope>NAPCQ</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>5PM</scope></search><sort><creationdate>200003</creationdate><title>Pharmacological characterization of small‐conductance Ca2+‐activated K+ channels stably expressed in HEK 293 cells</title><author>Strøbæk, Dorte ; Jørgensen, Tino D ; Christophersen, Palle ; Ahring, Philip K ; Olesen, Søren‐Peter</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-j3466-4267082adaf9b4857b1ba42adfaa9179f04a3df4f3d627ed2818f127de175c4b3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2000</creationdate><topic>apamin</topic><topic>bicuculline methiodide</topic><topic>Biological and medical sciences</topic><topic>Calcium‐activated potassium channel</topic><topic>Cell membranes. Ionic channels. Membrane pores</topic><topic>Cell structures and functions</topic><topic>dequalinium chloride</topic><topic>d‐tubocurarine</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>HEK 293</topic><topic>Molecular and cellular biology</topic><topic>scyllatoxin</topic><topic>taicatoxin</topic><topic>UCL 1684</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Strøbæk, Dorte</creatorcontrib><creatorcontrib>Jørgensen, Tino D</creatorcontrib><creatorcontrib>Christophersen, Palle</creatorcontrib><creatorcontrib>Ahring, Philip K</creatorcontrib><creatorcontrib>Olesen, Søren‐Peter</creatorcontrib><collection>Pascal-Francis</collection><collection>ProQuest Central (Corporate)</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Nursing & Allied Health Database</collection><collection>Neurosciences Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ProQuest Biological Science Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Biological Science Database</collection><collection>Nursing & Allied Health Premium</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>British journal of pharmacology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Strøbæk, Dorte</au><au>Jørgensen, Tino D</au><au>Christophersen, Palle</au><au>Ahring, Philip K</au><au>Olesen, Søren‐Peter</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Pharmacological characterization of small‐conductance Ca2+‐activated K+ channels stably expressed in HEK 293 cells</atitle><jtitle>British journal of pharmacology</jtitle><date>2000-03</date><risdate>2000</risdate><volume>129</volume><issue>5</issue><spage>991</spage><epage>999</epage><pages>991-999</pages><issn>0007-1188</issn><eissn>1476-5381</eissn><coden>BJPCBM</coden><abstract>Three genes encode the small‐conductance Ca2+‐activated K+ channels (SK channels). We have stably expressed hSK1 and rSK2 in HEK 293 cells and addressed the pharmacology of these subtypes using whole‐cell patch clamp recordings.
The bee venom peptide apamin blocked hSK1 as well as rSK2 with IC50 values of 3.3 nM and 83 pM, respectively.
The pharmacological separation between the subtypes was even more prominent when applying the scorpion peptide blocker scyllatoxin, which blocked hSK1 with an IC50 value of 80 nM and rSK2 at 287 pM.
The potent small molecule blockers showed little differentiation between the channel subtypes. The bis‐quinolinium cyclophane UCL 1684 blocked hSK1 with an IC50 value of 762 pM and rSK2 at 364 pM. The antiseptic compound dequalinium chloride blocked hSK1 and rSK2 with IC50 values of 444 nM and 162 nM, respectively.
The nicotinic acetylcholine receptor antagonist d‐tubocurarine was found to block hSK1 and rSK2 with IC50 values of 27 μM and 17 μM when measured at +80 mV. The inhibition by d‐tubocurarine was voltage‐dependent with increasing affinities at more hyperpolarized potentials.
The GABAA receptor antagonist bicuculline methiodide also blocked hSK1 and rSK2 in a voltage‐dependent manner with IC50 values of 15 and 25 μM when measured at +80 mV.
In conclusion, the pharmacological separation between SK channel subtypes expressed in mammalian cells is too small to support the notion that the apamin‐insensitive afterhyperpolarization of neurones is mediated by hSK1.
British Journal of Pharmacology (2000) 129, 991–999; doi:10.1038/sj.bjp.0703120</abstract><cop>Oxford, UK</cop><pub>Blackwell Publishing Ltd</pub><pmid>10696100</pmid><doi>10.1038/sj.bjp.0703120</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | apamin bicuculline methiodide Biological and medical sciences Calcium‐activated potassium channel Cell membranes. Ionic channels. Membrane pores Cell structures and functions dequalinium chloride d‐tubocurarine Fundamental and applied biological sciences. Psychology HEK 293 Molecular and cellular biology scyllatoxin taicatoxin UCL 1684 |
| title | Pharmacological characterization of small‐conductance Ca2+‐activated K+ channels stably expressed in HEK 293 cells |
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