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| <StructureSection load='6mz3' size='340' side='right'caption='[[6mz3]], [[Resolution|resolution]] 1.09Å' scene=''> | | <StructureSection load='6mz3' size='340' side='right'caption='[[6mz3]], [[Resolution|resolution]] 1.09Å' scene=''> |
| == Structural highlights == | | == Structural highlights == |
| <table><tr><td colspan='2'>[[6mz3]] is a 1 chain structure with sequence from [http://en.wikipedia.org/wiki/Dissp Dissp]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=6MZ3 OCA]. For a <b>guided tour on the structure components</b> use [http://proteopedia.org/fgij/fg.htm?mol=6MZ3 FirstGlance]. <br> | | <table><tr><td colspan='2'>[[6mz3]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Discosoma_sp. Discosoma sp.]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=6MZ3 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=6MZ3 FirstGlance]. <br> |
| </td></tr><tr id='NonStdRes'><td class="sblockLbl"><b>[[Non-Standard_Residue|NonStd Res:]]</b></td><td class="sblockDat"><scene name='pdbligand=CRO:{2-[(1R,2R)-1-AMINO-2-HYDROXYPROPYL]-4-(4-HYDROXYBENZYLIDENE)-5-OXO-4,5-DIHYDRO-1H-IMIDAZOL-1-YL}ACETIC+ACID'>CRO</scene></td></tr> | | </td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 1.088Å</td></tr> |
| <tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">PAmCherry, PAmCherry1 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=86600 DISSP])</td></tr>
| | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=CRO:{2-[(1R,2R)-1-AMINO-2-HYDROXYPROPYL]-4-(4-HYDROXYBENZYLIDENE)-5-OXO-4,5-DIHYDRO-1H-IMIDAZOL-1-YL}ACETIC+ACID'>CRO</scene></td></tr> |
| <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://proteopedia.org/fgij/fg.htm?mol=6mz3 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=6mz3 OCA], [http://pdbe.org/6mz3 PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=6mz3 RCSB], [http://www.ebi.ac.uk/pdbsum/6mz3 PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=6mz3 ProSAT]</span></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=6mz3 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=6mz3 OCA], [https://pdbe.org/6mz3 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=6mz3 RCSB], [https://www.ebi.ac.uk/pdbsum/6mz3 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=6mz3 ProSAT]</span></td></tr> |
| </table> | | </table> |
| <div style="background-color:#fffaf0;">
| | == Function == |
| == Publication Abstract from PubMed == | | [https://www.uniprot.org/uniprot/D1MPT3_DISSP D1MPT3_DISSP] |
| Intracellular pH plays a key role in physiology, and its measurement in living specimens remains a crucial task in biology. Fluorescent protein-based pH sensors have gained widespread use, but there is limited spectral diversity for multicolor detection, and it remains a challenge to measure absolute pH values. Here we demonstrate that mCherryTYG is an excellent fluorescence lifetime pH sensor that significantly expands the modalities available for pH quantification in live cells. We first report the 1.09 A X-ray crystal structure of mCherryTYG, exhibiting a fully matured chromophore. We next determine that it has an extraordinarily large dynamic range with a 2 ns lifetime change from pH 5.5 to 9.0. Critically, we find that the sensor maintains a p Ka of 6.8 independent of environment, whether as the purified protein in solution or expressed in live cells. Furthermore, the lifetime measurements are robustly independent of total fluorescence intensity and scatter. We demonstrate that mCherryTYG is a highly effective sensor using time-resolved fluorescence spectroscopy on live-cell suspensions, which has been previously overlooked as an easily accessible approach for quantifying intracellular pH. As a red fluorescent sensor, we also demonstrate that mCherryTYG is spectrally compatible with the ATeam sensor and EGFP for simultaneous dual-color measurements of intracellular pH, ATP, and extracellular pH. In a proof-of-concept, we quantify acute respiration-dependent pH homeostasis that exhibits a stoichiometric relationship with the ATP-generating capacity of the carbon fuel choice in E. coli. Broadly speaking, our work presents a previously unemployed methodology that will greatly facilitate continuous pH quantification.
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| Quantifying Acute Fuel and Respiration Dependent pH Homeostasis in Live Cells Using the mCherryTYG Mutant as a Fluorescence Lifetime Sensor.,Haynes EP, Rajendran M, Henning CK, Mishra A, Lyon AM, Tantama M Anal Chem. 2019 Jul 2;91(13):8466-8475. doi: 10.1021/acs.analchem.9b01562. Epub, 2019 Jun 19. PMID:31247720<ref>PMID:31247720</ref>
| | ==See Also== |
| | | *[[Green Fluorescent Protein 3D structures|Green Fluorescent Protein 3D structures]] |
| From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br>
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| </div>
| |
| <div class="pdbe-citations 6mz3" style="background-color:#fffaf0;"></div>
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| == References == | |
| <references/>
| |
| __TOC__ | | __TOC__ |
| </StructureSection> | | </StructureSection> |
| [[Category: Dissp]] | | [[Category: Discosoma sp]] |
| [[Category: Large Structures]] | | [[Category: Large Structures]] |
| [[Category: Haynes, E P]] | | [[Category: Haynes EP]] |
| [[Category: Tantama, M]] | | [[Category: Tantama M]] |
| [[Category: Beta barrel]]
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| [[Category: Biosensor]]
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| [[Category: Fluorescent protein]]
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| [[Category: Luminescent protein]]
| |