Ion transport activity and optogenetics capability of light-driven Na+-pump KR2
KR2 from marine bacteria Krokinobacter eikastus is a light-driven Na + pumping rhodopsin family (NaRs) member that actively transports Na + and/or H + depending on the ionic state. We here report electrophysiological studies on KR2 to address ion-transport properties under various electrochemical po...
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| creator | Hososhima, Shoko Kandori, Hideki Tsunoda, Satoshi P. |
| description | KR2 from marine bacteria
Krokinobacter eikastus
is a light-driven Na
+
pumping rhodopsin family (NaRs) member that actively transports Na
+
and/or H
+
depending on the ionic state. We here report electrophysiological studies on KR2 to address ion-transport properties under various electrochemical potentials of Δ[Na
+
], ΔpH, membrane voltage and light quality, because the contributions of these on the pumping activity were less understood so far. After transient expression of KR2 in mammalian cultured cells (ND7/23 cells), photocurrents were measured by whole-cell patch clamp under various intracellular Na
+
and pH conditions. When KR2 was continuously illuminated with LED light, two distinct time constants were obtained depending on the Na
+
concentration. KR2 exhibited slow ion transport (τ
off
of 28 ms) below 1.1 mM NaCl and rapid transport (τ
off
of 11 ms) above 11 mM NaCl. This indicates distinct transporting kinetics of H
+
and Na
+
. Photocurrent amplitude (current density) depends on the intracellular Na
+
concentration, as is expected for a Na
+
pump. The M-intermediate in the photocycle of KR2 could be transferred into the dark state without net ion transport by blue light illumination on top of green light. The M intermediate was stabilized by higher membrane voltage. Furthermore, we assessed the optogenetic silencing effect of rat cortical neurons after expressing KR2. Light power dependency revealed that action potential was profoundly inhibited by 1.5 mW/mm
2
green light illumination, confirming the ability to apply KR2 as an optogenetics silencer. |
| doi_str_mv | 10.1371/journal.pone.0256728 |
| format | Article |
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Krokinobacter eikastus
is a light-driven Na
+
pumping rhodopsin family (NaRs) member that actively transports Na
+
and/or H
+
depending on the ionic state. We here report electrophysiological studies on KR2 to address ion-transport properties under various electrochemical potentials of Δ[Na
+
], ΔpH, membrane voltage and light quality, because the contributions of these on the pumping activity were less understood so far. After transient expression of KR2 in mammalian cultured cells (ND7/23 cells), photocurrents were measured by whole-cell patch clamp under various intracellular Na
+
and pH conditions. When KR2 was continuously illuminated with LED light, two distinct time constants were obtained depending on the Na
+
concentration. KR2 exhibited slow ion transport (τ
off
of 28 ms) below 1.1 mM NaCl and rapid transport (τ
off
of 11 ms) above 11 mM NaCl. This indicates distinct transporting kinetics of H
+
and Na
+
. Photocurrent amplitude (current density) depends on the intracellular Na
+
concentration, as is expected for a Na
+
pump. The M-intermediate in the photocycle of KR2 could be transferred into the dark state without net ion transport by blue light illumination on top of green light. The M intermediate was stabilized by higher membrane voltage. Furthermore, we assessed the optogenetic silencing effect of rat cortical neurons after expressing KR2. Light power dependency revealed that action potential was profoundly inhibited by 1.5 mW/mm
2
green light illumination, confirming the ability to apply KR2 as an optogenetics silencer.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0256728</identifier><identifier>PMID: 34506508</identifier><language>eng</language><publisher>San Francisco: Public Library of Science</publisher><subject>Action potential ; Binding sites ; Biology and Life Sciences ; Electric potential ; Electrochemistry ; Engineering and Technology ; Genetics ; Hydrogen ; Illumination ; Information processing ; Intracellular ; Ion transport ; Life sciences ; Light ; Light emitting diodes ; Light quality ; Membranes ; Na+/H+-exchanging ATPase ; Optics ; Photoelectric effect ; Photoelectric emission ; Physical Sciences ; Plasmids ; Pumping ; Regulatory sequences ; Rhodopsin ; Sodium channels (voltage-gated) ; Sodium chloride ; Transport properties ; Voltage</subject><ispartof>PloS one, 2021-09, Vol.16 (9), p.e0256728-e0256728</ispartof><rights>2021 Hososhima et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2021 Hososhima et al 2021 Hososhima et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c569t-b5597239c94c481022394f10dcebbeed89cba1a089a1c775f25d4cd286d5651e3</citedby><cites>FETCH-LOGICAL-c569t-b5597239c94c481022394f10dcebbeed89cba1a089a1c775f25d4cd286d5651e3</cites><orcidid>0000-0003-3636-1521</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC8432791/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC8432791/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,864,885,2102,2928,23866,27924,27925,53791,53793</link.rule.ids></links><search><contributor>Obukhov, Alexander G</contributor><creatorcontrib>Hososhima, Shoko</creatorcontrib><creatorcontrib>Kandori, Hideki</creatorcontrib><creatorcontrib>Tsunoda, Satoshi P.</creatorcontrib><title>Ion transport activity and optogenetics capability of light-driven Na+-pump KR2</title><title>PloS one</title><description>KR2 from marine bacteria
Krokinobacter eikastus
is a light-driven Na
+
pumping rhodopsin family (NaRs) member that actively transports Na
+
and/or H
+
depending on the ionic state. We here report electrophysiological studies on KR2 to address ion-transport properties under various electrochemical potentials of Δ[Na
+
], ΔpH, membrane voltage and light quality, because the contributions of these on the pumping activity were less understood so far. After transient expression of KR2 in mammalian cultured cells (ND7/23 cells), photocurrents were measured by whole-cell patch clamp under various intracellular Na
+
and pH conditions. When KR2 was continuously illuminated with LED light, two distinct time constants were obtained depending on the Na
+
concentration. KR2 exhibited slow ion transport (τ
off
of 28 ms) below 1.1 mM NaCl and rapid transport (τ
off
of 11 ms) above 11 mM NaCl. This indicates distinct transporting kinetics of H
+
and Na
+
. Photocurrent amplitude (current density) depends on the intracellular Na
+
concentration, as is expected for a Na
+
pump. The M-intermediate in the photocycle of KR2 could be transferred into the dark state without net ion transport by blue light illumination on top of green light. The M intermediate was stabilized by higher membrane voltage. Furthermore, we assessed the optogenetic silencing effect of rat cortical neurons after expressing KR2. Light power dependency revealed that action potential was profoundly inhibited by 1.5 mW/mm
2
green light illumination, confirming the ability to apply KR2 as an optogenetics silencer.</description><subject>Action potential</subject><subject>Binding sites</subject><subject>Biology and Life Sciences</subject><subject>Electric potential</subject><subject>Electrochemistry</subject><subject>Engineering and Technology</subject><subject>Genetics</subject><subject>Hydrogen</subject><subject>Illumination</subject><subject>Information processing</subject><subject>Intracellular</subject><subject>Ion transport</subject><subject>Life sciences</subject><subject>Light</subject><subject>Light emitting diodes</subject><subject>Light quality</subject><subject>Membranes</subject><subject>Na+/H+-exchanging ATPase</subject><subject>Optics</subject><subject>Photoelectric effect</subject><subject>Photoelectric emission</subject><subject>Physical Sciences</subject><subject>Plasmids</subject><subject>Pumping</subject><subject>Regulatory sequences</subject><subject>Rhodopsin</subject><subject>Sodium channels (voltage-gated)</subject><subject>Sodium chloride</subject><subject>Transport 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transport activity and optogenetics capability of light-driven Na+-pump KR2</title><author>Hososhima, Shoko ; Kandori, Hideki ; Tsunoda, Satoshi P.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c569t-b5597239c94c481022394f10dcebbeed89cba1a089a1c775f25d4cd286d5651e3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2021</creationdate><topic>Action potential</topic><topic>Binding sites</topic><topic>Biology and Life Sciences</topic><topic>Electric potential</topic><topic>Electrochemistry</topic><topic>Engineering and Technology</topic><topic>Genetics</topic><topic>Hydrogen</topic><topic>Illumination</topic><topic>Information processing</topic><topic>Intracellular</topic><topic>Ion transport</topic><topic>Life sciences</topic><topic>Light</topic><topic>Light emitting diodes</topic><topic>Light quality</topic><topic>Membranes</topic><topic>Na+/H+-exchanging ATPase</topic><topic>Optics</topic><topic>Photoelectric effect</topic><topic>Photoelectric emission</topic><topic>Physical Sciences</topic><topic>Plasmids</topic><topic>Pumping</topic><topic>Regulatory sequences</topic><topic>Rhodopsin</topic><topic>Sodium channels (voltage-gated)</topic><topic>Sodium chloride</topic><topic>Transport properties</topic><topic>Voltage</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Hososhima, Shoko</creatorcontrib><creatorcontrib>Kandori, Hideki</creatorcontrib><creatorcontrib>Tsunoda, Satoshi P.</creatorcontrib><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Animal Behavior Abstracts</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Nursing & Allied Health Database</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full 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Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Hososhima, Shoko</au><au>Kandori, Hideki</au><au>Tsunoda, Satoshi P.</au><au>Obukhov, Alexander G</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Ion transport activity and optogenetics capability of light-driven Na+-pump KR2</atitle><jtitle>PloS one</jtitle><date>2021-09-10</date><risdate>2021</risdate><volume>16</volume><issue>9</issue><spage>e0256728</spage><epage>e0256728</epage><pages>e0256728-e0256728</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>KR2 from marine bacteria
Krokinobacter eikastus
is a light-driven Na
+
pumping rhodopsin family (NaRs) member that actively transports Na
+
and/or H
+
depending on the ionic state. We here report electrophysiological studies on KR2 to address ion-transport properties under various electrochemical potentials of Δ[Na
+
], ΔpH, membrane voltage and light quality, because the contributions of these on the pumping activity were less understood so far. After transient expression of KR2 in mammalian cultured cells (ND7/23 cells), photocurrents were measured by whole-cell patch clamp under various intracellular Na
+
and pH conditions. When KR2 was continuously illuminated with LED light, two distinct time constants were obtained depending on the Na
+
concentration. KR2 exhibited slow ion transport (τ
off
of 28 ms) below 1.1 mM NaCl and rapid transport (τ
off
of 11 ms) above 11 mM NaCl. This indicates distinct transporting kinetics of H
+
and Na
+
. Photocurrent amplitude (current density) depends on the intracellular Na
+
concentration, as is expected for a Na
+
pump. The M-intermediate in the photocycle of KR2 could be transferred into the dark state without net ion transport by blue light illumination on top of green light. The M intermediate was stabilized by higher membrane voltage. Furthermore, we assessed the optogenetic silencing effect of rat cortical neurons after expressing KR2. Light power dependency revealed that action potential was profoundly inhibited by 1.5 mW/mm
2
green light illumination, confirming the ability to apply KR2 as an optogenetics silencer.</abstract><cop>San Francisco</cop><pub>Public Library of Science</pub><pmid>34506508</pmid><doi>10.1371/journal.pone.0256728</doi><orcidid>https://orcid.org/0000-0003-3636-1521</orcidid><oa>free_for_read</oa></addata></record> |
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| language | eng |
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| source | DOAJ Directory of Open Access Journals; Public Library of Science (PLoS) Journals Open Access; EZB-FREE-00999 freely available EZB journals; PubMed Central; Free Full-Text Journals in Chemistry |
| subjects | Action potential Binding sites Biology and Life Sciences Electric potential Electrochemistry Engineering and Technology Genetics Hydrogen Illumination Information processing Intracellular Ion transport Life sciences Light Light emitting diodes Light quality Membranes Na+/H+-exchanging ATPase Optics Photoelectric effect Photoelectric emission Physical Sciences Plasmids Pumping Regulatory sequences Rhodopsin Sodium channels (voltage-gated) Sodium chloride Transport properties Voltage |
| title | Ion transport activity and optogenetics capability of light-driven Na+-pump KR2 |
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