6j1a: Difference between revisions
New page: '''Unreleased structure''' The entry 6j1a is ON HOLD Authors: Nakashima, R., Sakurai, K., shinoda, H., Matsuda, T., Nagai, T. Description: Photoswitchable fluorescent protein Gamillus,... |
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==Photoswitchable fluorescent protein Gamillus, off-state== | |||
<StructureSection load='6j1a' size='340' side='right'caption='[[6j1a]], [[Resolution|resolution]] 1.96Å' scene=''> | |||
== Structural highlights == | |||
<table><tr><td colspan='2'>[[6j1a]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Olindias Olindias]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=6J1A OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=6J1A FirstGlance]. <br> | |||
</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 1.96Å</td></tr> | |||
<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=CL:CHLORIDE+ION'>CL</scene>, <scene name='pdbligand=GOL:GLYCEROL'>GOL</scene>, <scene name='pdbligand=PO4:PHOSPHATE+ION'>PO4</scene>, <scene name='pdbligand=QYG:{(4E)-2-[(1S)-1,4-diamino-4-oxobutyl]-4-[(4-hydroxyphenyl)methylidene]-5-oxo-4,5-dihydro-1H-imidazol-1-yl}acetic+acid'>QYG</scene></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=6j1a FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=6j1a OCA], [https://pdbe.org/6j1a PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=6j1a RCSB], [https://www.ebi.ac.uk/pdbsum/6j1a PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=6j1a ProSAT]</span></td></tr> | |||
</table> | |||
== Function == | |||
[https://www.uniprot.org/uniprot/A0A2Z5X7U3_9CNID A0A2Z5X7U3_9CNID] | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
Reversibly switchable fluorescent proteins (RSFPs) are crucial tags for super-resolution observation of protein localization and dynamics inside living cells. However, due to the high fluorescence pKa ( approximately 5-6) of most RSFPs, their usage in acidic conditions (pH 4.5-6.0) has been limited. Here, we investigated a new photochromic mechanism in Gamillus, a recently developed green fluorescent protein with acid tolerance. Gamillus exhibits negative switching with especially high contrast in acidic conditions, and its off switching is caused by trans-to-cis isomerization of the chromophore hydroxyphenyl ring that accompanies protonation. Through a combination of rational design and saturation mutagenesis, we developed two variants with enhanced switching contrasts and off-switching speeds, designated rsGamillus-S and rsGamillus-F, respectively. The fluorescence intensity, off-switching speed, and switching contrast of the rsGamillus variants are only slightly affected by changes in pH between 4.5 and 7.5. Exploiting these properties, we succeeded in high-contrast super-resolution imaging of cellular architectures in acidic conditions. | |||
Acid-Tolerant Reversibly Switchable Green Fluorescent Protein for Super-resolution Imaging under Acidic Conditions.,Shinoda H, Lu K, Nakashima R, Wazawa T, Noguchi K, Matsuda T, Nagai T Cell Chem Biol. 2019 Oct 17;26(10):1469-1479.e6. doi:, 10.1016/j.chembiol.2019.07.012. Epub 2019 Aug 15. PMID:31422907<ref>PMID:31422907</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
[[Category: | </div> | ||
[[Category: | <div class="pdbe-citations 6j1a" style="background-color:#fffaf0;"></div> | ||
[[Category: | |||
[[Category: | ==See Also== | ||
[[Category: Nakashima | *[[Green Fluorescent Protein 3D structures|Green Fluorescent Protein 3D structures]] | ||
[[Category: | == References == | ||
<references/> | |||
__TOC__ | |||
</StructureSection> | |||
[[Category: Large Structures]] | |||
[[Category: Olindias]] | |||
[[Category: Matsuda T]] | |||
[[Category: Nagai T]] | |||
[[Category: Nakashima R]] | |||
[[Category: Sakurai K]] | |||
[[Category: Shinoda H]] | |||
Latest revision as of 13:26, 15 November 2023
Photoswitchable fluorescent protein Gamillus, off-statePhotoswitchable fluorescent protein Gamillus, off-state
Structural highlights
FunctionPublication Abstract from PubMedReversibly switchable fluorescent proteins (RSFPs) are crucial tags for super-resolution observation of protein localization and dynamics inside living cells. However, due to the high fluorescence pKa ( approximately 5-6) of most RSFPs, their usage in acidic conditions (pH 4.5-6.0) has been limited. Here, we investigated a new photochromic mechanism in Gamillus, a recently developed green fluorescent protein with acid tolerance. Gamillus exhibits negative switching with especially high contrast in acidic conditions, and its off switching is caused by trans-to-cis isomerization of the chromophore hydroxyphenyl ring that accompanies protonation. Through a combination of rational design and saturation mutagenesis, we developed two variants with enhanced switching contrasts and off-switching speeds, designated rsGamillus-S and rsGamillus-F, respectively. The fluorescence intensity, off-switching speed, and switching contrast of the rsGamillus variants are only slightly affected by changes in pH between 4.5 and 7.5. Exploiting these properties, we succeeded in high-contrast super-resolution imaging of cellular architectures in acidic conditions. Acid-Tolerant Reversibly Switchable Green Fluorescent Protein for Super-resolution Imaging under Acidic Conditions.,Shinoda H, Lu K, Nakashima R, Wazawa T, Noguchi K, Matsuda T, Nagai T Cell Chem Biol. 2019 Oct 17;26(10):1469-1479.e6. doi:, 10.1016/j.chembiol.2019.07.012. Epub 2019 Aug 15. PMID:31422907[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. See AlsoReferences
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