4r6b: Difference between revisions
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==Rational Design of Enhanced Photoresistance in a Photoswitchable Fluorescent Protein== | ==Rational Design of Enhanced Photoresistance in a Photoswitchable Fluorescent Protein== | ||
<StructureSection load='4r6b' size='340' side='right' caption='[[4r6b]], [[Resolution|resolution]] 2.00Å' scene=''> | <StructureSection load='4r6b' size='340' side='right'caption='[[4r6b]], [[Resolution|resolution]] 2.00Å' scene=''> | ||
== Structural highlights == | == Structural highlights == | ||
<table><tr><td colspan='2'>[[4r6b]] is a 4 chain structure with sequence from [ | <table><tr><td colspan='2'>[[4r6b]] is a 4 chain structure with sequence from [https://en.wikipedia.org/wiki/Lobophyllia_hemprichii Lobophyllia hemprichii]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4R6B OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=4R6B FirstGlance]. <br> | ||
</td></tr><tr id=' | </td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 2Å</td></tr> | ||
<tr id=' | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=CR8:2-[1-AMINO-2-(1H-IMIDAZOL-5-YL)ETHYL]-1-(CARBOXYMETHYL)-4-[(4-OXOCYCLOHEXA-2,5-DIEN-1-YLIDENE)METHYL]-1H-IMIDAZOL-5-OLATE'>CR8</scene>, <scene name='pdbligand=SO3:SULFITE+ION'>SO3</scene>, <scene name='pdbligand=SO4:SULFATE+ION'>SO4</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=4r6b FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4r6b OCA], [https://pdbe.org/4r6b PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=4r6b RCSB], [https://www.ebi.ac.uk/pdbsum/4r6b PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=4r6b ProSAT]</span></td></tr> | ||
<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[ | |||
</table> | </table> | ||
== Function == | |||
[https://www.uniprot.org/uniprot/Q5S6Z9_LOBHE Q5S6Z9_LOBHE] | |||
<div style="background-color:#fffaf0;"> | <div style="background-color:#fffaf0;"> | ||
== Publication Abstract from PubMed == | == Publication Abstract from PubMed == | ||
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__TOC__ | __TOC__ | ||
</StructureSection> | </StructureSection> | ||
[[Category: | [[Category: Large Structures]] | ||
[[Category: Adam | [[Category: Lobophyllia hemprichii]] | ||
[[Category: Bourgeois | [[Category: Adam V]] | ||
[[Category: Byrdin | [[Category: Bourgeois D]] | ||
[[Category: Duan | [[Category: Byrdin M]] | ||
[[Category: Duan C]] | |||
Latest revision as of 20:44, 20 September 2023
Rational Design of Enhanced Photoresistance in a Photoswitchable Fluorescent ProteinRational Design of Enhanced Photoresistance in a Photoswitchable Fluorescent Protein
Structural highlights
FunctionPublication Abstract from PubMedFluorescent proteins are particularly susceptible to photobleaching, the permanent loss of fluorescence emission resulting from photodestruction of the chromophore. In the case of Reversibly Switchable Fluorescent Proteins (RSFPs), which can be switched back and forth between a non-fluorescent and a fluorescent state, the achievable number of switching cycles is limited by photobleaching, a process known as photofatigue. Photofatigue has become a crucial limitation in a number of advanced applications based on repeated photoswitching of RSFPs, notably in the field of super-resolution fluorescence microscopy. Here, based on our previous structural investigation of photobleaching mechanisms in IrisFP, an RSFP also capable of green-to-red photoconversion, we present the rational design of a single-mutant IrisFP-M159A that displays considerably enhanced photostability. The results suggest that, under moderate illumination intensities, photobleaching of IrisFP-like Anthozoan fluorescent proteins such as EosFP, Dendra or Dronpa derivatives is mainly driven by an oxygen-dependent mechanism resulting in the irreversible sulfoxidation of methionine 159. The photofatigue decay profiles of IrisFP and its photoresistant mutant IrisFP-M159A were investigated in different experimental conditions, in vitro and in cellulo. Although the performance of the mutant was found to be always superior, the results showed switching behaviors strongly dependent on the nanoenvironment. Thus, in general, assessment of photostability and switching properties of RSFPs should be carried out in real experimental conditions. Rational design of enhanced photoresistance in a photoswitchable fluorescent protein.,Duan C, Byrdin M, El Khatib M, Henry X, Adam V, Bourgeois D Methods Appl Fluoresc. 2015 Jan 16;3(1):014004. doi:, 10.1088/2050-6120/3/1/014004. PMID:29148481[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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