Structural Determinants of the High Affinity Extracellular Zinc Binding Site on Cav3.2 T-type Calcium Channels
Cav3.2 T-type channels contain a high affinity metal binding site for trace metals such as copper and zinc. This site is occupied at physiologically relevant concentrations of these metals, leading to decreased channel activity and pain transmission. A histidine at position 191 was recently identifi...
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Veröffentlicht in: | The Journal of biological chemistry 2010-01, Vol.285 (5), p.3271-3281 |
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description | Cav3.2 T-type channels contain a high affinity metal binding site for trace metals such as copper and zinc. This site is occupied at physiologically relevant concentrations of these metals, leading to decreased channel activity and pain transmission. A histidine at position 191 was recently identified as a critical determinant for both trace metal block of Cav3.2 and modulation by redox agents. His191 is found on the extracellular face of the Cav3.2 channel on the IS3-S4 linker and is not conserved in other Cav3 channels. Mutation of the corresponding residue in Cav3.1 to histidine, Gln172, significantly enhances trace metal inhibition, but not to the level observed in wild-type Cav3.2, implying that other residues also contribute to the metal binding site. The goal of the present study is to identify these other residues using a series of chimeric channels. The key findings of the study are that the metal binding site is composed of a Asp-Gly-His motif in IS3–S4 and a second aspartate residue in IS2. These results suggest that metal binding stabilizes the closed conformation of the voltage-sensor paddle in repeat I, and thereby inhibits channel opening. These studies provide insight into the structure of T-type channels, and identify an extracellular motif that could be targeted for drug development. |
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This site is occupied at physiologically relevant concentrations of these metals, leading to decreased channel activity and pain transmission. A histidine at position 191 was recently identified as a critical determinant for both trace metal block of Cav3.2 and modulation by redox agents. His191 is found on the extracellular face of the Cav3.2 channel on the IS3-S4 linker and is not conserved in other Cav3 channels. Mutation of the corresponding residue in Cav3.1 to histidine, Gln172, significantly enhances trace metal inhibition, but not to the level observed in wild-type Cav3.2, implying that other residues also contribute to the metal binding site. The goal of the present study is to identify these other residues using a series of chimeric channels. The key findings of the study are that the metal binding site is composed of a Asp-Gly-His motif in IS3–S4 and a second aspartate residue in IS2. These results suggest that metal binding stabilizes the closed conformation of the voltage-sensor paddle in repeat I, and thereby inhibits channel opening. These studies provide insight into the structure of T-type channels, and identify an extracellular motif that could be targeted for drug development.</description><identifier>ISSN: 0021-9258</identifier><identifier>EISSN: 1083-351X</identifier><identifier>DOI: 10.1074/jbc.M109.067660</identifier><identifier>PMID: 19940152</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Animals ; Aspartic Acid - chemistry ; Binding Sites ; Calcium Channels, T-Type - chemistry ; Cav3.2 ; Channels/Calcium ; Electrophysiology ; Electrophysiology - methods ; Female ; Histidine - chemistry ; Inhibitory Concentration 50 ; Membrane Transport, Structure, Function, and Biogenesis ; Membrane/Channels ; Metals/Zinc ; Models, Molecular ; Mutation ; Oocytes - metabolism ; Oxidation-Reduction ; Rats ; T-type Calcium Channel ; Voltage-clamping ; Xenopus ; Xenopus Oocytes ; Zinc - chemistry</subject><ispartof>The Journal of biological chemistry, 2010-01, Vol.285 (5), p.3271-3281</ispartof><rights>2010 © 2010 ASBMB. Currently published by Elsevier Inc; originally published by American Society for Biochemistry and Molecular Biology.</rights><rights>2010 by The American Society for Biochemistry and Molecular Biology, Inc.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4080-45c50a5966e7c7f70174648bb9b81a85ec4f385415761a582f93a4de95700d923</citedby><cites>FETCH-LOGICAL-c4080-45c50a5966e7c7f70174648bb9b81a85ec4f385415761a582f93a4de95700d923</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC2823403/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC2823403/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,315,728,781,785,886,27928,27929,53795,53797</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/19940152$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Kang, Ho-Won</creatorcontrib><creatorcontrib>Vitko, Iuliia</creatorcontrib><creatorcontrib>Lee, Sang-Soo</creatorcontrib><creatorcontrib>Perez-Reyes, Edward</creatorcontrib><creatorcontrib>Lee, Jung-Ha</creatorcontrib><title>Structural Determinants of the High Affinity Extracellular Zinc Binding Site on Cav3.2 T-type Calcium Channels</title><title>The Journal of biological chemistry</title><addtitle>J Biol Chem</addtitle><description>Cav3.2 T-type channels contain a high affinity metal binding site for trace metals such as copper and zinc. This site is occupied at physiologically relevant concentrations of these metals, leading to decreased channel activity and pain transmission. A histidine at position 191 was recently identified as a critical determinant for both trace metal block of Cav3.2 and modulation by redox agents. His191 is found on the extracellular face of the Cav3.2 channel on the IS3-S4 linker and is not conserved in other Cav3 channels. Mutation of the corresponding residue in Cav3.1 to histidine, Gln172, significantly enhances trace metal inhibition, but not to the level observed in wild-type Cav3.2, implying that other residues also contribute to the metal binding site. The goal of the present study is to identify these other residues using a series of chimeric channels. The key findings of the study are that the metal binding site is composed of a Asp-Gly-His motif in IS3–S4 and a second aspartate residue in IS2. These results suggest that metal binding stabilizes the closed conformation of the voltage-sensor paddle in repeat I, and thereby inhibits channel opening. These studies provide insight into the structure of T-type channels, and identify an extracellular motif that could be targeted for drug development.</description><subject>Animals</subject><subject>Aspartic Acid - chemistry</subject><subject>Binding Sites</subject><subject>Calcium Channels, T-Type - chemistry</subject><subject>Cav3.2</subject><subject>Channels/Calcium</subject><subject>Electrophysiology</subject><subject>Electrophysiology - methods</subject><subject>Female</subject><subject>Histidine - chemistry</subject><subject>Inhibitory Concentration 50</subject><subject>Membrane Transport, Structure, Function, and Biogenesis</subject><subject>Membrane/Channels</subject><subject>Metals/Zinc</subject><subject>Models, Molecular</subject><subject>Mutation</subject><subject>Oocytes - metabolism</subject><subject>Oxidation-Reduction</subject><subject>Rats</subject><subject>T-type Calcium Channel</subject><subject>Voltage-clamping</subject><subject>Xenopus</subject><subject>Xenopus Oocytes</subject><subject>Zinc - chemistry</subject><issn>0021-9258</issn><issn>1083-351X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kEtv1DAURi0EotPCmh1Y7DP1M443SGUoFKmIxbQSYmM5zs3EVeKMHGfo_HtcUvFY4I1l-dzvfjoIvaJkTYkS53e1W3-hRK9JqcqSPEErSipecEm_PUUrQhgtNJPVCTqdpjuSj9D0OTqhWgtCJVuhsE1xdmmOtscfIEEcfLAhTXhsceoAX_ldhy_a1gefjvjyPkXroO_n3kb83QeH3_vQ-LDDW58AjwFv7IGvGb4p0nEP-dU7Pw9409kQoJ9eoGet7Sd4-XifoduPlzebq-L666fPm4vrwglSkUJIJ4mVuixBOdUqQpUoRVXXuq6orSQ40fJKCipVSa2sWKu5FQ1oqQhpNONn6N2Su5_rARoHIRfvzT76wcajGa03__4E35ndeDCsYlwQngPOlwAXx2mK0P6epcQ8qDdZvXlQbxb1eeL13yv_8I-uM_B2Abrs9IePYGo_ug6GvFQaaThTNENvFqi1o7G76Cdzu2WE8qxAC_qrmV6IrBMOHqKZnIfgoMmRLplm9P_t-BPYB6el</recordid><startdate>20100129</startdate><enddate>20100129</enddate><creator>Kang, Ho-Won</creator><creator>Vitko, Iuliia</creator><creator>Lee, Sang-Soo</creator><creator>Perez-Reyes, Edward</creator><creator>Lee, Jung-Ha</creator><general>Elsevier Inc</general><general>American Society for Biochemistry and Molecular Biology</general><scope>6I.</scope><scope>AAFTH</scope><scope>FBQ</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>5PM</scope></search><sort><creationdate>20100129</creationdate><title>Structural Determinants of the High Affinity Extracellular Zinc Binding Site on Cav3.2 T-type Calcium Channels</title><author>Kang, Ho-Won ; Vitko, Iuliia ; Lee, Sang-Soo ; Perez-Reyes, Edward ; Lee, Jung-Ha</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4080-45c50a5966e7c7f70174648bb9b81a85ec4f385415761a582f93a4de95700d923</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2010</creationdate><topic>Animals</topic><topic>Aspartic Acid - chemistry</topic><topic>Binding Sites</topic><topic>Calcium Channels, T-Type - chemistry</topic><topic>Cav3.2</topic><topic>Channels/Calcium</topic><topic>Electrophysiology</topic><topic>Electrophysiology - methods</topic><topic>Female</topic><topic>Histidine - chemistry</topic><topic>Inhibitory Concentration 50</topic><topic>Membrane Transport, Structure, Function, and Biogenesis</topic><topic>Membrane/Channels</topic><topic>Metals/Zinc</topic><topic>Models, Molecular</topic><topic>Mutation</topic><topic>Oocytes - metabolism</topic><topic>Oxidation-Reduction</topic><topic>Rats</topic><topic>T-type Calcium Channel</topic><topic>Voltage-clamping</topic><topic>Xenopus</topic><topic>Xenopus Oocytes</topic><topic>Zinc - chemistry</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kang, Ho-Won</creatorcontrib><creatorcontrib>Vitko, Iuliia</creatorcontrib><creatorcontrib>Lee, Sang-Soo</creatorcontrib><creatorcontrib>Perez-Reyes, Edward</creatorcontrib><creatorcontrib>Lee, Jung-Ha</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>AGRIS</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>The Journal of biological chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kang, Ho-Won</au><au>Vitko, Iuliia</au><au>Lee, Sang-Soo</au><au>Perez-Reyes, Edward</au><au>Lee, Jung-Ha</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Structural Determinants of the High Affinity Extracellular Zinc Binding Site on Cav3.2 T-type Calcium Channels</atitle><jtitle>The Journal of biological chemistry</jtitle><addtitle>J Biol Chem</addtitle><date>2010-01-29</date><risdate>2010</risdate><volume>285</volume><issue>5</issue><spage>3271</spage><epage>3281</epage><pages>3271-3281</pages><issn>0021-9258</issn><eissn>1083-351X</eissn><abstract>Cav3.2 T-type channels contain a high affinity metal binding site for trace metals such as copper and zinc. This site is occupied at physiologically relevant concentrations of these metals, leading to decreased channel activity and pain transmission. A histidine at position 191 was recently identified as a critical determinant for both trace metal block of Cav3.2 and modulation by redox agents. His191 is found on the extracellular face of the Cav3.2 channel on the IS3-S4 linker and is not conserved in other Cav3 channels. Mutation of the corresponding residue in Cav3.1 to histidine, Gln172, significantly enhances trace metal inhibition, but not to the level observed in wild-type Cav3.2, implying that other residues also contribute to the metal binding site. The goal of the present study is to identify these other residues using a series of chimeric channels. The key findings of the study are that the metal binding site is composed of a Asp-Gly-His motif in IS3–S4 and a second aspartate residue in IS2. These results suggest that metal binding stabilizes the closed conformation of the voltage-sensor paddle in repeat I, and thereby inhibits channel opening. These studies provide insight into the structure of T-type channels, and identify an extracellular motif that could be targeted for drug development.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>19940152</pmid><doi>10.1074/jbc.M109.067660</doi><tpages>11</tpages><oa>free_for_read</oa></addata></record> |
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subjects | Animals Aspartic Acid - chemistry Binding Sites Calcium Channels, T-Type - chemistry Cav3.2 Channels/Calcium Electrophysiology Electrophysiology - methods Female Histidine - chemistry Inhibitory Concentration 50 Membrane Transport, Structure, Function, and Biogenesis Membrane/Channels Metals/Zinc Models, Molecular Mutation Oocytes - metabolism Oxidation-Reduction Rats T-type Calcium Channel Voltage-clamping Xenopus Xenopus Oocytes Zinc - chemistry |
title | Structural Determinants of the High Affinity Extracellular Zinc Binding Site on Cav3.2 T-type Calcium Channels |
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