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Kinetics of proton release and uptake by channelrhodopsin-2
► Transient proton translocation by channelrhodopsin-2 was detected by an optical pH indicator. ► Proton release and uptake proceed with time constants of 1.7ms and 11ms, respectively. ► The response time of the pH indicator is not limited by surface-to-bulk diffusion. ► Residues E123 and S245 do no...
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Published in: | FEBS letters 2012-05, Vol.586 (9), p.1344-1348 |
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creator | Nack, Melanie Radu, Ionela Schultz, Bernd-Joachim Resler, Tom Schlesinger, Ramona Bondar, Ana-Nicoleta del Val, Coral Abbruzzetti, Stefania Viappiani, Cristiano Bamann, Christian Bamberg, Ernst Heberle, Joachim |
description | ► Transient proton translocation by channelrhodopsin-2 was detected by an optical pH indicator. ► Proton release and uptake proceed with time constants of 1.7ms and 11ms, respectively. ► The response time of the pH indicator is not limited by surface-to-bulk diffusion. ► Residues E123 and S245 do not participate in proton release. ► The concurrence of proton release and ion channeling points to a mechanistic link.
Electrophysiological experiments showed that the light-activated cation channel channelrhodopsin-2 (ChR2) pumps protons in the absence of a membrane potential. We determined here the kinetics of transient pH change using a water-soluble pH-indicator. It is shown that ChR2 released protons prior to uptake with a stoichiometry of 0.3 protons per ChR2. Comparison to the photocycle kinetics revealed that proton release and uptake match rise and decay of the P3520 intermediate. As the P3520 state also represents the conductive state of cation channeling, the concurrence of proton pumping and channel gating implies an intimate mechanistic link of the two functional modes. Studies on the E123T and S245E mutants show that these residues are not critically involved in proton translocation. |
doi_str_mv | 10.1016/j.febslet.2012.03.047 |
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Electrophysiological experiments showed that the light-activated cation channel channelrhodopsin-2 (ChR2) pumps protons in the absence of a membrane potential. We determined here the kinetics of transient pH change using a water-soluble pH-indicator. It is shown that ChR2 released protons prior to uptake with a stoichiometry of 0.3 protons per ChR2. Comparison to the photocycle kinetics revealed that proton release and uptake match rise and decay of the P3520 intermediate. As the P3520 state also represents the conductive state of cation channeling, the concurrence of proton pumping and channel gating implies an intimate mechanistic link of the two functional modes. Studies on the E123T and S245E mutants show that these residues are not critically involved in proton translocation.</description><identifier>ISSN: 0014-5793</identifier><identifier>EISSN: 1873-3468</identifier><identifier>DOI: 10.1016/j.febslet.2012.03.047</identifier><identifier>PMID: 22504075</identifier><language>eng</language><publisher>England: Elsevier B.V</publisher><subject>Bacteriorhodopsin ; Biological Transport ; Hydrogen-Ion Concentration ; Ion channel ; Kinetics ; Models, Molecular ; Optogenetics ; Protein Structure, Secondary ; Proton Pumps - chemistry ; Proton Pumps - metabolism ; Proton transfer ; Protons ; Rhodopsin ; Rhodopsin - chemistry ; Rhodopsin - metabolism</subject><ispartof>FEBS letters, 2012-05, Vol.586 (9), p.1344-1348</ispartof><rights>2012 Federation of European Biochemical Societies</rights><rights>FEBS Letters 586 (2012) 1873-3468 © 2015 Federation of European Biochemical Societies</rights><rights>Copyright © 2012 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4965-94fde2b8a7e3931eaf2e56b274e36b51641017ffee15ba054243191e2b32aec23</citedby><cites>FETCH-LOGICAL-c4965-94fde2b8a7e3931eaf2e56b274e36b51641017ffee15ba054243191e2b32aec23</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0014579312002475$$EHTML$$P50$$Gelsevier$$Hfree_for_read</linktohtml><link.rule.ids>314,777,781,3536,27905,27906,45761</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/22504075$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Nack, Melanie</creatorcontrib><creatorcontrib>Radu, Ionela</creatorcontrib><creatorcontrib>Schultz, Bernd-Joachim</creatorcontrib><creatorcontrib>Resler, Tom</creatorcontrib><creatorcontrib>Schlesinger, Ramona</creatorcontrib><creatorcontrib>Bondar, Ana-Nicoleta</creatorcontrib><creatorcontrib>del Val, Coral</creatorcontrib><creatorcontrib>Abbruzzetti, Stefania</creatorcontrib><creatorcontrib>Viappiani, Cristiano</creatorcontrib><creatorcontrib>Bamann, Christian</creatorcontrib><creatorcontrib>Bamberg, Ernst</creatorcontrib><creatorcontrib>Heberle, Joachim</creatorcontrib><title>Kinetics of proton release and uptake by channelrhodopsin-2</title><title>FEBS letters</title><addtitle>FEBS Lett</addtitle><description>► Transient proton translocation by channelrhodopsin-2 was detected by an optical pH indicator. ► Proton release and uptake proceed with time constants of 1.7ms and 11ms, respectively. ► The response time of the pH indicator is not limited by surface-to-bulk diffusion. ► Residues E123 and S245 do not participate in proton release. ► The concurrence of proton release and ion channeling points to a mechanistic link.
Electrophysiological experiments showed that the light-activated cation channel channelrhodopsin-2 (ChR2) pumps protons in the absence of a membrane potential. We determined here the kinetics of transient pH change using a water-soluble pH-indicator. It is shown that ChR2 released protons prior to uptake with a stoichiometry of 0.3 protons per ChR2. Comparison to the photocycle kinetics revealed that proton release and uptake match rise and decay of the P3520 intermediate. As the P3520 state also represents the conductive state of cation channeling, the concurrence of proton pumping and channel gating implies an intimate mechanistic link of the two functional modes. Studies on the E123T and S245E mutants show that these residues are not critically involved in proton translocation.</description><subject>Bacteriorhodopsin</subject><subject>Biological Transport</subject><subject>Hydrogen-Ion Concentration</subject><subject>Ion channel</subject><subject>Kinetics</subject><subject>Models, Molecular</subject><subject>Optogenetics</subject><subject>Protein Structure, Secondary</subject><subject>Proton Pumps - chemistry</subject><subject>Proton Pumps - metabolism</subject><subject>Proton transfer</subject><subject>Protons</subject><subject>Rhodopsin</subject><subject>Rhodopsin - chemistry</subject><subject>Rhodopsin - metabolism</subject><issn>0014-5793</issn><issn>1873-3468</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2012</creationdate><recordtype>article</recordtype><recordid>eNqNkctOwzAQRS0EoqXwCaAs2ST4mYdYIKhaiqjEAlhbjjNRXdKk2Amof49DCltYWZbOvR6fQeic4IhgEl-toxJyV0EbUUxohFmEeXKAxiRNWMh4nB6iMcaEhyLJ2AidOLfG_p6S7BiNKBWY40SM0fWjqaE12gVNGWxt0zZ1YKEC5SBQdRF021a9QZDvAr1SdQ2VXTVFs3WmDukpOipV5eBsf07Q63z2Ml2Ey6f7h-ntMtQ8i0WY8bIAmqcqAZYxAqqkIOKcJhxYnAsSc_-hpCwBiMgVFpxyRjLiI4wq0JRN0OXQ6-d778C1cmOchqpSNTSdkz6OeSoSknpUDKi2jXMWSrm1ZqPszkM9F8u13HuTvTeJmfTefO5i_0SXb6D4Tf2I8sBiAD5NBbv_tcr57I4-90vod0AoxpR_V90MVeCdfRiw0mkDtYbCWNCtLBrzx7RfA1SVcA</recordid><startdate>20120507</startdate><enddate>20120507</enddate><creator>Nack, Melanie</creator><creator>Radu, Ionela</creator><creator>Schultz, Bernd-Joachim</creator><creator>Resler, Tom</creator><creator>Schlesinger, Ramona</creator><creator>Bondar, Ana-Nicoleta</creator><creator>del Val, Coral</creator><creator>Abbruzzetti, Stefania</creator><creator>Viappiani, Cristiano</creator><creator>Bamann, Christian</creator><creator>Bamberg, Ernst</creator><creator>Heberle, Joachim</creator><general>Elsevier B.V</general><scope>6I.</scope><scope>AAFTH</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>7X8</scope></search><sort><creationdate>20120507</creationdate><title>Kinetics of proton release and uptake by channelrhodopsin-2</title><author>Nack, Melanie ; Radu, Ionela ; Schultz, Bernd-Joachim ; Resler, Tom ; Schlesinger, Ramona ; Bondar, Ana-Nicoleta ; del Val, Coral ; Abbruzzetti, Stefania ; Viappiani, Cristiano ; Bamann, Christian ; Bamberg, Ernst ; Heberle, Joachim</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4965-94fde2b8a7e3931eaf2e56b274e36b51641017ffee15ba054243191e2b32aec23</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2012</creationdate><topic>Bacteriorhodopsin</topic><topic>Biological Transport</topic><topic>Hydrogen-Ion Concentration</topic><topic>Ion channel</topic><topic>Kinetics</topic><topic>Models, Molecular</topic><topic>Optogenetics</topic><topic>Protein Structure, Secondary</topic><topic>Proton Pumps - chemistry</topic><topic>Proton Pumps - metabolism</topic><topic>Proton transfer</topic><topic>Protons</topic><topic>Rhodopsin</topic><topic>Rhodopsin - chemistry</topic><topic>Rhodopsin - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Nack, Melanie</creatorcontrib><creatorcontrib>Radu, Ionela</creatorcontrib><creatorcontrib>Schultz, Bernd-Joachim</creatorcontrib><creatorcontrib>Resler, Tom</creatorcontrib><creatorcontrib>Schlesinger, Ramona</creatorcontrib><creatorcontrib>Bondar, Ana-Nicoleta</creatorcontrib><creatorcontrib>del Val, Coral</creatorcontrib><creatorcontrib>Abbruzzetti, Stefania</creatorcontrib><creatorcontrib>Viappiani, Cristiano</creatorcontrib><creatorcontrib>Bamann, Christian</creatorcontrib><creatorcontrib>Bamberg, Ernst</creatorcontrib><creatorcontrib>Heberle, Joachim</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>FEBS letters</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Nack, Melanie</au><au>Radu, Ionela</au><au>Schultz, Bernd-Joachim</au><au>Resler, Tom</au><au>Schlesinger, Ramona</au><au>Bondar, Ana-Nicoleta</au><au>del Val, Coral</au><au>Abbruzzetti, Stefania</au><au>Viappiani, Cristiano</au><au>Bamann, Christian</au><au>Bamberg, Ernst</au><au>Heberle, Joachim</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Kinetics of proton release and uptake by channelrhodopsin-2</atitle><jtitle>FEBS letters</jtitle><addtitle>FEBS Lett</addtitle><date>2012-05-07</date><risdate>2012</risdate><volume>586</volume><issue>9</issue><spage>1344</spage><epage>1348</epage><pages>1344-1348</pages><issn>0014-5793</issn><eissn>1873-3468</eissn><abstract>► Transient proton translocation by channelrhodopsin-2 was detected by an optical pH indicator. ► Proton release and uptake proceed with time constants of 1.7ms and 11ms, respectively. ► The response time of the pH indicator is not limited by surface-to-bulk diffusion. ► Residues E123 and S245 do not participate in proton release. ► The concurrence of proton release and ion channeling points to a mechanistic link.
Electrophysiological experiments showed that the light-activated cation channel channelrhodopsin-2 (ChR2) pumps protons in the absence of a membrane potential. We determined here the kinetics of transient pH change using a water-soluble pH-indicator. It is shown that ChR2 released protons prior to uptake with a stoichiometry of 0.3 protons per ChR2. Comparison to the photocycle kinetics revealed that proton release and uptake match rise and decay of the P3520 intermediate. As the P3520 state also represents the conductive state of cation channeling, the concurrence of proton pumping and channel gating implies an intimate mechanistic link of the two functional modes. Studies on the E123T and S245E mutants show that these residues are not critically involved in proton translocation.</abstract><cop>England</cop><pub>Elsevier B.V</pub><pmid>22504075</pmid><doi>10.1016/j.febslet.2012.03.047</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
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subjects | Bacteriorhodopsin Biological Transport Hydrogen-Ion Concentration Ion channel Kinetics Models, Molecular Optogenetics Protein Structure, Secondary Proton Pumps - chemistry Proton Pumps - metabolism Proton transfer Protons Rhodopsin Rhodopsin - chemistry Rhodopsin - metabolism |
title | Kinetics of proton release and uptake by channelrhodopsin-2 |
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