The Expanding Family of Natural Anion Channelrhodopsins Reveals Large Variations in Kinetics, Conductance, and Spectral Sensitivity
Identifieur interne : 000205 ( Pmc/Curation ); précédent : 000204; suivant : 000206The Expanding Family of Natural Anion Channelrhodopsins Reveals Large Variations in Kinetics, Conductance, and Spectral Sensitivity
Auteurs : Elena G. Govorunova [États-Unis] ; Oleg A. Sineshchekov [États-Unis] ; Elsa M. Rodarte [États-Unis] ; Roger Janz [États-Unis] ; Olivier Morelle [Allemagne] ; Michael Melkonian [Allemagne] ; Gane K.-S. Wong [Canada, République populaire de Chine] ; John L. Spudich [États-Unis]Source :
- Scientific Reports [ 2045-2322 ] ; 2017.
Abstract
Natural anion channelrhodopsins (ACRs) discovered in the cryptophyte alga
Url:
DOI: 10.1038/srep43358
PubMed: 28256618
PubMed Central: 5335703
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">The Expanding Family of Natural Anion Channelrhodopsins Reveals Large Variations in Kinetics, Conductance, and Spectral Sensitivity</title><author><name sortKey="Govorunova, Elena G" sort="Govorunova, Elena G" uniqKey="Govorunova E" first="Elena G." last="Govorunova">Elena G. Govorunova</name><affiliation wicri:level="1"><nlm:aff id="a1"><institution>Center for Membrane Biology, Department of Biochemistry & Molecular Biology, The University of Texas Health Science Center at Houston, McGovern Medical School</institution>, Houston, Texas,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Sineshchekov, Oleg A" sort="Sineshchekov, Oleg A" uniqKey="Sineshchekov O" first="Oleg A." last="Sineshchekov">Oleg A. Sineshchekov</name><affiliation wicri:level="1"><nlm:aff id="a1"><institution>Center for Membrane Biology, Department of Biochemistry & Molecular Biology, The University of Texas Health Science Center at Houston, McGovern Medical School</institution>, Houston, Texas,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Rodarte, Elsa M" sort="Rodarte, Elsa M" uniqKey="Rodarte E" first="Elsa M." last="Rodarte">Elsa M. Rodarte</name><affiliation wicri:level="1"><nlm:aff id="a2"><institution>Department of Neurobiology & Anatomy, The University of Texas Health Science Center at Houston, McGovern Medical School</institution>, Houston, Texas,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Janz, Roger" sort="Janz, Roger" uniqKey="Janz R" first="Roger" last="Janz">Roger Janz</name><affiliation wicri:level="1"><nlm:aff id="a2"><institution>Department of Neurobiology & Anatomy, The University of Texas Health Science Center at Houston, McGovern Medical School</institution>, Houston, Texas,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Morelle, Olivier" sort="Morelle, Olivier" uniqKey="Morelle O" first="Olivier" last="Morelle">Olivier Morelle</name><affiliation wicri:level="1"><nlm:aff id="a3"><institution>Institute of Botany, Cologne Biocenter, University of Cologne</institution>, Cologne,<country>Germany</country></nlm:aff><country xml:lang="fr">Allemagne</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Melkonian, Michael" sort="Melkonian, Michael" uniqKey="Melkonian M" first="Michael" last="Melkonian">Michael Melkonian</name><affiliation wicri:level="1"><nlm:aff id="a3"><institution>Institute of Botany, Cologne Biocenter, University of Cologne</institution>, Cologne,<country>Germany</country></nlm:aff><country xml:lang="fr">Allemagne</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Wong, Gane K S" sort="Wong, Gane K S" uniqKey="Wong G" first="Gane K.-S." last="Wong">Gane K.-S. Wong</name><affiliation wicri:level="1"><nlm:aff id="a4"><institution>Departments of Biological Sciences and of Medicine, University of Alberta, Edmonton</institution>, Alberta,<country>Canada</country></nlm:aff><country xml:lang="fr">Canada</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="a5"><institution>BGI-Shenzhen</institution>, Shenzhen,<country>China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Spudich, John L" sort="Spudich, John L" uniqKey="Spudich J" first="John L." last="Spudich">John L. Spudich</name><affiliation wicri:level="1"><nlm:aff id="a1"><institution>Center for Membrane Biology, Department of Biochemistry & Molecular Biology, The University of Texas Health Science Center at Houston, McGovern Medical School</institution>, Houston, Texas,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">28256618</idno><idno type="pmc">5335703</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5335703</idno><idno type="RBID">PMC:5335703</idno><idno type="doi">10.1038/srep43358</idno><date when="2017">2017</date><idno type="wicri:Area/Pmc/Corpus">000205</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000205</idno><idno type="wicri:Area/Pmc/Curation">000205</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">000205</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">The Expanding Family of Natural Anion Channelrhodopsins Reveals Large Variations in Kinetics, Conductance, and Spectral Sensitivity</title><author><name sortKey="Govorunova, Elena G" sort="Govorunova, Elena G" uniqKey="Govorunova E" first="Elena G." last="Govorunova">Elena G. Govorunova</name><affiliation wicri:level="1"><nlm:aff id="a1"><institution>Center for Membrane Biology, Department of Biochemistry & Molecular Biology, The University of Texas Health Science Center at Houston, McGovern Medical School</institution>, Houston, Texas,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Sineshchekov, Oleg A" sort="Sineshchekov, Oleg A" uniqKey="Sineshchekov O" first="Oleg A." last="Sineshchekov">Oleg A. Sineshchekov</name><affiliation wicri:level="1"><nlm:aff id="a1"><institution>Center for Membrane Biology, Department of Biochemistry & Molecular Biology, The University of Texas Health Science Center at Houston, McGovern Medical School</institution>, Houston, Texas,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Rodarte, Elsa M" sort="Rodarte, Elsa M" uniqKey="Rodarte E" first="Elsa M." last="Rodarte">Elsa M. Rodarte</name><affiliation wicri:level="1"><nlm:aff id="a2"><institution>Department of Neurobiology & Anatomy, The University of Texas Health Science Center at Houston, McGovern Medical School</institution>, Houston, Texas,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Janz, Roger" sort="Janz, Roger" uniqKey="Janz R" first="Roger" last="Janz">Roger Janz</name><affiliation wicri:level="1"><nlm:aff id="a2"><institution>Department of Neurobiology & Anatomy, The University of Texas Health Science Center at Houston, McGovern Medical School</institution>, Houston, Texas,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Morelle, Olivier" sort="Morelle, Olivier" uniqKey="Morelle O" first="Olivier" last="Morelle">Olivier Morelle</name><affiliation wicri:level="1"><nlm:aff id="a3"><institution>Institute of Botany, Cologne Biocenter, University of Cologne</institution>, Cologne,<country>Germany</country></nlm:aff><country xml:lang="fr">Allemagne</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Melkonian, Michael" sort="Melkonian, Michael" uniqKey="Melkonian M" first="Michael" last="Melkonian">Michael Melkonian</name><affiliation wicri:level="1"><nlm:aff id="a3"><institution>Institute of Botany, Cologne Biocenter, University of Cologne</institution>, Cologne,<country>Germany</country></nlm:aff><country xml:lang="fr">Allemagne</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Wong, Gane K S" sort="Wong, Gane K S" uniqKey="Wong G" first="Gane K.-S." last="Wong">Gane K.-S. Wong</name><affiliation wicri:level="1"><nlm:aff id="a4"><institution>Departments of Biological Sciences and of Medicine, University of Alberta, Edmonton</institution>, Alberta,<country>Canada</country></nlm:aff><country xml:lang="fr">Canada</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="a5"><institution>BGI-Shenzhen</institution>, Shenzhen,<country>China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Spudich, John L" sort="Spudich, John L" uniqKey="Spudich J" first="John L." last="Spudich">John L. Spudich</name><affiliation wicri:level="1"><nlm:aff id="a1"><institution>Center for Membrane Biology, Department of Biochemistry & Molecular Biology, The University of Texas Health Science Center at Houston, McGovern Medical School</institution>, Houston, Texas,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author></analytic><series><title level="j">Scientific Reports</title><idno type="eISSN">2045-2322</idno><imprint><date when="2017">2017</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Natural anion channelrhodopsins (ACRs) discovered in the cryptophyte alga <italic>Guillardia theta</italic> generate large hyperpolarizing currents at membrane potentials above the Nernst equilibrium potential for Cl<sup>−</sup> and thus can be used as efficient inhibitory tools for optogenetics. We have identified and characterized new ACR homologs in different cryptophyte species, showing that all of them are anion-selective, and thus expanded this protein family to 20 functionally confirmed members. Sequence comparison of natural ACRs and engineered Cl<sup>−</sup>-conducting mutants of cation channelrhodopsins (CCRs) showed radical differences in their anion selectivity filters. In particular, the Glu90 residue in channelrhodopsin 2, which needed to be mutated to a neutral or alkaline residue to confer anion selectivity to CCRs, is nevertheless conserved in all of the ACRs identified. The new ACRs showed a large variation of the amplitude, kinetics, and spectral sensitivity of their photocurrents. A notable variant, designated “ZipACR”, is particularly promising for inhibitory optogenetics because of its combination of larger current amplitudes than those of previously reported ACRs and an unprecedentedly fast conductance cycle (current half-decay time 2–4 ms depending on voltage). ZipACR expressed in cultured mouse hippocampal neurons enabled precise photoinhibition of individual spikes in trains of up to 50 Hz frequency.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Deisseroth, K" uniqKey="Deisseroth K">K. Deisseroth</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Zhang, F" uniqKey="Zhang F">F. Zhang</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Chow, B Y" uniqKey="Chow B">B. Y. Chow</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Maurice, N" uniqKey="Maurice N">N. 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<front><journal-meta><journal-id journal-id-type="nlm-ta">Sci Rep</journal-id><journal-id journal-id-type="iso-abbrev">Sci Rep</journal-id><journal-title-group><journal-title>Scientific Reports</journal-title></journal-title-group><issn pub-type="epub">2045-2322</issn><publisher><publisher-name>Nature Publishing Group</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">28256618</article-id><article-id pub-id-type="pmc">5335703</article-id><article-id pub-id-type="pii">srep43358</article-id><article-id pub-id-type="doi">10.1038/srep43358</article-id><article-categories><subj-group subj-group-type="heading"><subject>Article</subject></subj-group></article-categories><title-group><article-title>The Expanding Family of Natural Anion Channelrhodopsins Reveals Large Variations in Kinetics, Conductance, and Spectral Sensitivity</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Govorunova</surname><given-names>Elena G.</given-names></name><xref ref-type="aff" rid="a1">1</xref></contrib><contrib contrib-type="author"><name><surname>Sineshchekov</surname><given-names>Oleg A.</given-names></name><xref ref-type="aff" rid="a1">1</xref></contrib><contrib contrib-type="author"><name><surname>Rodarte</surname><given-names>Elsa M.</given-names></name><xref ref-type="aff" rid="a2">2</xref></contrib><contrib contrib-type="author"><name><surname>Janz</surname><given-names>Roger</given-names></name><xref ref-type="aff" rid="a2">2</xref></contrib><contrib contrib-type="author"><name><surname>Morelle</surname><given-names>Olivier</given-names></name><xref ref-type="aff" rid="a3">3</xref></contrib><contrib contrib-type="author"><name><surname>Melkonian</surname><given-names>Michael</given-names></name><xref ref-type="aff" rid="a3">3</xref></contrib><contrib contrib-type="author"><name><surname>Wong</surname><given-names>Gane K.-S.</given-names></name><xref ref-type="aff" rid="a4">4</xref><xref ref-type="aff" rid="a5">5</xref></contrib><contrib contrib-type="author"><name><surname>Spudich</surname><given-names>John L.</given-names></name><xref ref-type="corresp" rid="c1">a</xref><xref ref-type="aff" rid="a1">1</xref></contrib><aff id="a1"><label>1</label><institution>Center for Membrane Biology, Department of Biochemistry & Molecular Biology, The University of Texas Health Science Center at Houston, McGovern Medical School</institution>, Houston, Texas,<country>USA</country></aff><aff id="a2"><label>2</label><institution>Department of Neurobiology & Anatomy, The University of Texas Health Science Center at Houston, McGovern Medical School</institution>, Houston, Texas,<country>USA</country></aff><aff id="a3"><label>3</label><institution>Institute of Botany, Cologne Biocenter, University of Cologne</institution>, Cologne,<country>Germany</country></aff><aff id="a4"><label>4</label><institution>Departments of Biological Sciences and of Medicine, University of Alberta, Edmonton</institution>, Alberta,<country>Canada</country></aff><aff id="a5"><label>5</label><institution>BGI-Shenzhen</institution>, Shenzhen,<country>China</country></aff></contrib-group><author-notes><corresp id="c1"><label>a</label><email>John.L.Spudich@uth.tmc.edu</email></corresp></author-notes><pub-date pub-type="epub"><day>03</day><month>03</month><year>2017</year></pub-date><pub-date pub-type="collection"><year>2017</year></pub-date><volume>7</volume><elocation-id>43358</elocation-id><history><date date-type="received"><day>05</day><month>10</month><year>2016</year></date><date date-type="accepted"><day>23</day><month>01</month><year>2017</year></date></history><permissions><copyright-statement>Copyright © 2017, The Author(s)</copyright-statement><copyright-year>2017</copyright-year><copyright-holder>The Author(s)</copyright-holder><license license-type="open-access" xlink:href="http://creativecommons.org/licenses/by/4.0/"><pmc-comment>author-paid</pmc-comment>
<license-p>This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit <ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/licenses/by/4.0/">http://creativecommons.org/licenses/by/4.0/</ext-link></license-p></license></permissions><abstract><p>Natural anion channelrhodopsins (ACRs) discovered in the cryptophyte alga <italic>Guillardia theta</italic> generate large hyperpolarizing currents at membrane potentials above the Nernst equilibrium potential for Cl<sup>−</sup> and thus can be used as efficient inhibitory tools for optogenetics. We have identified and characterized new ACR homologs in different cryptophyte species, showing that all of them are anion-selective, and thus expanded this protein family to 20 functionally confirmed members. Sequence comparison of natural ACRs and engineered Cl<sup>−</sup>-conducting mutants of cation channelrhodopsins (CCRs) showed radical differences in their anion selectivity filters. In particular, the Glu90 residue in channelrhodopsin 2, which needed to be mutated to a neutral or alkaline residue to confer anion selectivity to CCRs, is nevertheless conserved in all of the ACRs identified. The new ACRs showed a large variation of the amplitude, kinetics, and spectral sensitivity of their photocurrents. A notable variant, designated “ZipACR”, is particularly promising for inhibitory optogenetics because of its combination of larger current amplitudes than those of previously reported ACRs and an unprecedentedly fast conductance cycle (current half-decay time 2–4 ms depending on voltage). ZipACR expressed in cultured mouse hippocampal neurons enabled precise photoinhibition of individual spikes in trains of up to 50 Hz frequency.</p></abstract></article-meta></front><floats-group><fig id="f1"><label>Figure 1</label><caption><title>Action spectra of photocurrents generated in HEK293 cells.</title><p>An example of a spectrally shifted ACR pair from a single cryptophyte species (<italic>C1</italic>ACR_023 and <italic>C1</italic>ACR_887), and an example of three spectrally matching ACRs from another species (<italic>R1</italic>ACR_367, <italic>R1</italic>ACR_877 and <italic>R1</italic>ACR_741). The initial slopes of photocurrents were measured in the linear range of the intensity dependence, corrected for the quantum density, and normalized to the maximal value obtained for each protein (for more detail see Methods). The data points are the mean values ± sem (n = 4–6 measurements in 3–4 cells). The spectra of other proteins were measured in a similar way and the resultant spectral maxima are listed in <xref ref-type="table" rid="t1">Table 1</xref>.</p></caption><graphic xlink:href="srep43358-f1"></graphic></fig><fig id="f2"><label>Figure 2</label><caption><title>All functional ACR homologs conduct Cl<sup>−</sup>.</title><p>(<bold>a</bold>) A representative series of photocurrent traces recorded in response to a 1-s light pulse of the saturating intensity from <italic>Psu</italic>ACR<italic>_</italic>973 (ZipACR) expressed in HEK293 cells in the standard bath (black lines) and upon partial replacement of Cl<sup>−</sup> with Asp<sup>−</sup> in the bath (red lines). The holding voltage (E<sub>h</sub>) was changed in 20-mV steps from −60 mV at the amplifier output (bottom trace). (<bold>b</bold>) The current-voltage relationships of peak (filled symbols) and stationary (empty symbols) photocurrents determined from current traces as shown in panel a. The data points are the mean values ± sem (n = 3 measurements in the same typical cell). The data were corrected for liquid junction potentials. (<bold>c</bold>) The E<sub>r</sub> shifts (E<sub>r</sub> in the standard bath minus E<sub>r</sub> in the Asp bath) measured for the currents averaged over the entire 1-s illumination period for all tested ACR homologs and <italic>Gt</italic>ACR1 mutants. The data are mean values ± sd (n = 3–5 cells). The data for the earlier studied ACRs are included for comparison.</p></caption><graphic xlink:href="srep43358-f2"></graphic></fig><fig id="f3"><label>Figure 3</label><caption><title>Comparison of the mutations introduced to confer Cl<sup>−</sup> selectivity to CCRs with the corresponding residues in natural ACRs.</title><p>The color code is: red, negatively charged residues; blue, positively charged residues; green, polar residues; yellow, non-polar residues. The numbers on top show the residue numbers according to the <italic>Cr</italic>ChR2 sequence.</p></caption><graphic xlink:href="srep43358-f3"></graphic></fig><fig id="f4"><label>Figure 4</label><caption><title>ACR functional diversity.</title><p>The peak current amplitude in response to a 1-s light pulse of the saturating intensity, decay rate (measured as the reciprocal of the time of the 50% decrease in the current amplitude after the light-off) and wavelength of maximal sensitivity (determined by continuous approximation of experimental data points as shown in <xref ref-type="fig" rid="f1">Fig. 1</xref>) of cryptophyte ACR homologs. The amplitude and decay rate data were obtained at −60 mV holding potential in standard solutions (see Methods). The data points are the mean values; for sem values and the number of sampled cells see <xref ref-type="supplementary-material" rid="S1">Supplementary Table S2</xref>. The numbers next to the data points correspond to the protein numbers in <xref ref-type="table" rid="t1">Table 1</xref>. The data for the previously known <italic>Gt</italic>ACR1, <italic>Gt</italic>ACR2 and <italic>Psu</italic>ACR1 are included for comparison.</p></caption><graphic xlink:href="srep43358-f4"></graphic></fig><fig id="f5"><label>Figure 5</label><caption><title>ZipACR is a fast inhibitory tool for optogenetics.</title><p>(<bold>a</bold>) The decay of photocurrents generated by ACRs indicated in the figure legend in HEK293 cells after a 1-s pulse of continuous light of the saturating intensity at the wavelength of the peak absorption of the respective pigment. The traces were normalized at the stationary level measured near the end of the illumination period. (<bold>b</bold>) Photoinduced spike suppression in cultured mouse hippocampal neurons expressing ZipACR (red line) or <italic>Gt</italic>ACR2 (black line). Neurons were stimulated by injection of 1-ms current pulses at 50 Hz (schematically shown below). The wavelength of the 20-ms light pulses of the saturating intensity (shown on top of each trace) was 520 nm for ZipACR and 470 nm for <italic>Gt</italic>ACR2. (<bold>c</bold>) Photoinduced changes in the normalized spike amplitude measured as shown in panel b. The data points are the mean values ± sem (n = 7 cells). The amplitude was normalized to the mean amplitude of the last five spikes before switching on the light.</p></caption><graphic xlink:href="srep43358-f5"></graphic></fig><table-wrap position="float" id="t1"><label>Table 1</label><caption><title>GenBank accession numbers, source organisms, transcript names, action spectra maxima and protein name abbreviations of ACR homologs tested in this study.</title></caption><table frame="hsides" rules="groups" border="1"><colgroup><col align="left"></col><col align="center"></col><col align="center"></col><col align="center"></col><col align="center"></col><col align="center"></col></colgroup><thead valign="bottom"><tr><th align="left" valign="top" charoff="50">#</th><th align="center" valign="top" charoff="50">Accession</th><th align="center" valign="top" charoff="50">Organism</th><th align="center" valign="top" charoff="50">Transcript name</th><th align="center" valign="top" charoff="50">Spectral max. (nm)</th><th align="center" valign="top" charoff="50">Protein name abbreviation</th></tr></thead><tbody valign="top"><tr><td align="left" valign="top" charoff="50">1</td><td align="center" valign="top" charoff="50">KX879674</td><td rowspan="3" align="center" valign="middle" charoff="50"><italic>Geminigera cryophila</italic> (CCMP2564)</td><td align="center" valign="top" charoff="50">CAMNT 0021181457<xref ref-type="fn" rid="t1-fn1">*</xref></td><td align="center" valign="top" charoff="50">495</td><td align="center" valign="top" charoff="50"><italic>Gc</italic>ACR_457</td></tr><tr><td align="left" valign="top" charoff="50">2</td><td align="center" valign="top" charoff="50">KX879675</td><td align="center" valign="top" charoff="50">CAMNT 0021207145<xref ref-type="fn" rid="t1-fn1">*</xref></td><td align="center" valign="top" charoff="50">485</td><td align="center" valign="top" charoff="50"><italic>Gc</italic>ACR_145</td></tr><tr><td align="left" valign="top" charoff="50">3</td><td align="center" valign="top" charoff="50">KX879676</td><td align="center" valign="top" charoff="50">CAMNT 0021218439<xref ref-type="fn" rid="t1-fn1">*</xref></td><td align="center" valign="top" charoff="50">515</td><td align="center" valign="top" charoff="50"><italic>Gc</italic>ACR_439</td></tr><tr><td align="left" valign="top" charoff="50">4</td><td align="center" valign="top" charoff="50">KX879677</td><td rowspan="2" align="center" valign="middle" charoff="50"><italic>Geminigera</italic> sp. (Caron Lab Isolate)</td><td align="center" valign="top" charoff="50">CAMNT 0013945203<xref ref-type="fn" rid="t1-fn1">*</xref></td><td align="center" valign="top" charoff="50">475</td><td align="center" valign="top" charoff="50"><italic>G1</italic>ACR_203</td></tr><tr><td align="left" valign="top" charoff="50">5</td><td align="center" valign="top" charoff="50">KX879678</td><td align="center" valign="top" charoff="50">CAMNT 0013979243<xref ref-type="fn" rid="t1-fn1">*</xref></td><td align="center" valign="top" charoff="50">N/A</td><td align="center" valign="top" charoff="50"><italic>G1</italic>ACR<italic>_</italic>243</td></tr><tr><td align="left" valign="top" charoff="50">6</td><td align="center" valign="top" charoff="50">KP171708</td><td rowspan="2" align="center" valign="middle" charoff="50"><italic>Guillardia theta</italic> (CCMP2712)</td><td align="center" valign="top" charoff="50">known previously<xref ref-type="bibr" rid="b6">6</xref></td><td align="center" valign="top" charoff="50">515</td><td align="center" valign="top" charoff="50"><italic>Gt</italic>ACR1</td></tr><tr><td align="left" valign="top" charoff="50">7</td><td align="center" valign="top" charoff="50">KP171709</td><td align="center" valign="top" charoff="50">known previously<xref ref-type="bibr" rid="b6">6</xref></td><td align="center" valign="top" charoff="50">470</td><td align="center" valign="top" charoff="50"><italic>Gt</italic>ACR2</td></tr><tr><td align="left" valign="top" charoff="50">8</td><td align="center" valign="top" charoff="50">KF992074</td><td rowspan="4" align="center" valign="middle" charoff="50"><italic>Proteomonas sulcata</italic> (CCMP704)</td><td align="center" valign="top" charoff="50">known previously<xref ref-type="bibr" rid="b7">7</xref></td><td align="center" valign="top" charoff="50">520</td><td align="center" valign="top" charoff="50"><italic>Psu</italic>ACR1</td></tr><tr><td align="left" valign="top" charoff="50">9</td><td align="center" valign="top" charoff="50">KX879679</td><td align="center" valign="top" charoff="50">CAMNT 0026648973<xref ref-type="fn" rid="t1-fn1">*</xref></td><td align="center" valign="top" charoff="50">515</td><td align="center" valign="top" charoff="50"><italic>Psu</italic> ACR_973 (ZipACR)</td></tr><tr><td align="left" valign="top" charoff="50">10</td><td align="center" valign="top" charoff="50">KX879680</td><td align="center" valign="top" charoff="50">CAMNT 0026650433<xref ref-type="fn" rid="t1-fn1">*</xref></td><td align="center" valign="top" charoff="50">525</td><td align="center" valign="top" charoff="50"><italic>Psu</italic>ACR_433</td></tr><tr><td align="left" valign="top" charoff="50">11</td><td align="center" valign="top" charoff="50">KX879681</td><td align="center" valign="top" charoff="50">IRZA-2061003<xref ref-type="fn" rid="t1-fn2">#</xref></td><td align="center" valign="top" charoff="50">N/A</td><td align="center" valign="top" charoff="50"><italic>Psu</italic>ACR_003</td></tr><tr><td align="left" valign="top" charoff="50">12</td><td align="center" valign="top" charoff="50">KX879682</td><td align="center" valign="top" charoff="50"><italic>Rhodomonas lens</italic> (RHODO)</td><td align="center" valign="top" charoff="50">CAMNT 0019228477<xref ref-type="fn" rid="t1-fn1">*</xref></td><td align="center" valign="top" charoff="50">445</td><td align="center" valign="top" charoff="50"><italic>Rl</italic>ACR_477</td></tr><tr><td align="left" valign="top" charoff="50">13</td><td align="center" valign="top" charoff="50">KX879683</td><td align="center" valign="top" charoff="50"><italic>Rhodomonas salina</italic> (CCMP1319)</td><td align="center" valign="top" charoff="50">CAMNT 0012794995<xref ref-type="fn" rid="t1-fn1">*</xref></td><td align="center" valign="top" charoff="50">515</td><td align="center" valign="top" charoff="50"><italic>Rs</italic>ACR_995</td></tr><tr><td align="left" valign="top" charoff="50">14</td><td align="center" valign="top" charoff="50">KX879684</td><td rowspan="5" align="center" valign="middle" charoff="50"><italic>Rhodomonas</italic> sp. (CCMP768)</td><td align="center" valign="top" charoff="50">CAMNT 0042060367<xref ref-type="fn" rid="t1-fn1">*</xref></td><td align="center" valign="top" charoff="50">515</td><td align="center" valign="top" charoff="50"><italic>R1</italic>ACR_367</td></tr><tr><td align="left" valign="top" charoff="50">15</td><td align="center" valign="top" charoff="50">KX879685</td><td align="center" valign="top" charoff="50">CAMNT 0042061877<xref ref-type="fn" rid="t1-fn1">*</xref></td><td align="center" valign="top" charoff="50">520</td><td align="center" valign="top" charoff="50"><italic>R1</italic>ACR_877</td></tr><tr><td align="left" valign="top" charoff="50">16</td><td align="center" valign="top" charoff="50">KX879686</td><td align="center" valign="top" charoff="50">CAMNT 0042066447<xref ref-type="fn" rid="t1-fn1">*</xref></td><td align="center" valign="top" charoff="50">515</td><td align="center" valign="top" charoff="50"><italic>R1</italic>ACR_447</td></tr><tr><td align="left" valign="top" charoff="50">17</td><td align="center" valign="top" charoff="50">KX879687</td><td align="center" valign="top" charoff="50">CAMNT 0049477741<xref ref-type="fn" rid="t1-fn1">*</xref></td><td align="center" valign="top" charoff="50">515</td><td align="center" valign="top" charoff="50"><italic>R1</italic>ACR_741</td></tr><tr><td align="left" valign="top" charoff="50">18</td><td align="center" valign="top" charoff="50">KX879688</td><td align="center" valign="top" charoff="50">CAMNT 0049533799<xref ref-type="fn" rid="t1-fn1">*</xref></td><td align="center" valign="top" charoff="50">520</td><td align="center" valign="top" charoff="50"><italic>R1</italic>ACR_799</td></tr><tr><td align="left" valign="top" charoff="50">19</td><td align="center" valign="top" charoff="50">KX879689</td><td rowspan="2" align="center" valign="middle" charoff="50"><italic>Rhodomonas</italic> sp. (CCAC1630)</td><td align="center" valign="top" charoff="50">IAYV-2004853<xref ref-type="fn" rid="t1-fn2">#</xref></td><td align="center" valign="top" charoff="50">515</td><td align="center" valign="top" charoff="50"><italic>R2</italic>ACR_853</td></tr><tr><td align="left" valign="top" charoff="50">20</td><td align="center" valign="top" charoff="50">KX879690</td><td align="center" valign="top" charoff="50">IAYV-2007142<xref ref-type="fn" rid="t1-fn2">#</xref></td><td align="center" valign="top" charoff="50">490</td><td align="center" valign="top" charoff="50"><italic>R2</italic>ACR_142</td></tr><tr><td align="left" valign="top" charoff="50">21</td><td align="center" valign="top" charoff="50">KX879691</td><td rowspan="2" align="center" valign="middle" charoff="50">not classified (CCMP2293)</td><td align="center" valign="top" charoff="50">CAMNT 0022112887<xref ref-type="fn" rid="t1-fn1">*</xref></td><td align="center" valign="top" charoff="50">535</td><td align="center" valign="top" charoff="50"><italic>C1</italic>ACR_887</td></tr><tr><td align="left" valign="top" charoff="50">22</td><td align="center" valign="top" charoff="50">KX879692</td><td align="center" valign="top" charoff="50">CAMNT 0022176023<xref ref-type="fn" rid="t1-fn1">*</xref></td><td align="center" valign="top" charoff="50">445</td><td align="center" valign="top" charoff="50"><italic>C1</italic>ACR_023</td></tr></tbody></table><table-wrap-foot><fn id="t1-fn1"><p>Three previously known ACRs are also included.</p></fn><fn id="t1-fn2"><p><sup>*</sup>Transcripts from the MMETS project.</p></fn><fn id="t1-fn3"><p><sup>#</sup>Transcripts from the 1 KP project.</p></fn></table-wrap-foot></table-wrap></floats-group></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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