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| ==Optimized Ratiometric Calcium Sensors For Functional In Vivo Imaging of Neurons and T-Lymphocytes== | | ==Optimized Ratiometric Calcium Sensors For Functional In Vivo Imaging of Neurons and T-Lymphocytes== |
| <StructureSection load='2m97' size='340' side='right' caption='[[2m97]], [[NMR_Ensembles_of_Models | 20 NMR models]]' scene=''> | | <StructureSection load='2m97' size='340' side='right'caption='[[2m97]]' scene=''> |
| == Structural highlights == | | == Structural highlights == |
| <table><tr><td colspan='2'>[[2m97]] is a 1 chain structure with sequence from [http://en.wikipedia.org/wiki/Gadus_tau Gadus tau]. Full experimental information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2M97 OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=2M97 FirstGlance]. <br> | | <table><tr><td colspan='2'>[[2m97]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Opsanus_tau Opsanus tau]. Full experimental information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2M97 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=2M97 FirstGlance]. <br> |
| </td></tr><tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=2m97 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=2m97 OCA], [http://www.rcsb.org/pdb/explore.do?structureId=2m97 RCSB], [http://www.ebi.ac.uk/pdbsum/2m97 PDBsum]</span></td></tr> | | </td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">Solution NMR</td></tr> |
| | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=2m97 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=2m97 OCA], [https://pdbe.org/2m97 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=2m97 RCSB], [https://www.ebi.ac.uk/pdbsum/2m97 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=2m97 ProSAT]</span></td></tr> |
| </table> | | </table> |
| <div style="background-color:#fffaf0;">
| | == Function == |
| == Publication Abstract from PubMed == | | [https://www.uniprot.org/uniprot/W5IDB2_OPSTA W5IDB2_OPSTA] |
| The quality of genetically encoded calcium indicators (GECIs) has improved dramatically in recent years, but high-performing ratiometric indicators are still rare. Here we describe a series of fluorescence resonance energy transfer (FRET)-based calcium biosensors with a reduced number of calcium binding sites per sensor. These 'Twitch' sensors are based on the C-terminal domain of Opsanus troponin C. Their FRET responses were optimized by a large-scale functional screen in bacterial colonies, refined by a secondary screen in rat hippocampal neuron cultures. We tested the in vivo performance of the most sensitive variants in the brain and lymph nodes of mice. The sensitivity of the Twitch sensors matched that of synthetic calcium dyes and allowed visualization of tonic action potential firing in neurons and high resolution functional tracking of T lymphocytes. Given their ratiometric readout, their brightness, large dynamic range and linear response properties, Twitch sensors represent versatile tools for neuroscience and immunology.
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| Optimized ratiometric calcium sensors for functional in vivo imaging of neurons and T lymphocytes.,Thestrup T, Litzlbauer J, Bartholomaus I, Mues M, Russo L, Dana H, Kovalchuk Y, Liang Y, Kalamakis G, Laukat Y, Becker S, Witte G, Geiger A, Allen T, Rome LC, Chen TW, Kim DS, Garaschuk O, Griesinger C, Griesbeck O Nat Methods. 2014 Feb;11(2):175-82. doi: 10.1038/nmeth.2773. Epub 2014 Jan 5. PMID:24390440<ref>PMID:24390440</ref>
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| From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br>
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| </div>
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| == References ==
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| <references/>
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| __TOC__ | | __TOC__ |
| </StructureSection> | | </StructureSection> |
| [[Category: Gadus tau]] | | [[Category: Large Structures]] |
| [[Category: Becker, S]] | | [[Category: Opsanus tau]] |
| [[Category: Griesinger, C]] | | [[Category: Becker S]] |
| [[Category: Russo, L]] | | [[Category: Griesinger C]] |
| [[Category: Calcium biosensor]]
| | [[Category: Russo L]] |
| [[Category: Metal binding protein]]
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