2ye0: Difference between revisions
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==X-ray structure of the cyan fluorescent protein mTurquoise (K206A mutant)== | |||
<StructureSection load='2ye0' size='340' side='right'caption='[[2ye0]], [[Resolution|resolution]] 1.47Å' scene=''> | |||
== Structural highlights == | |||
<table><tr><td colspan='2'>[[2ye0]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Aequorea_victoria Aequorea victoria]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2YE0 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=2YE0 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]] 1.47Å</td></tr> | |||
<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=SWG:2-[(4Z)-2-[(1R)-1-AMINO-2-HYDROXY-ETHYL]-4-(1H-INDOL-3-YLMETHYLIDENE)-5-OXO-IMIDAZOL-1-YL]ETHANOIC+ACID'>SWG</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=2ye0 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=2ye0 OCA], [https://pdbe.org/2ye0 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=2ye0 RCSB], [https://www.ebi.ac.uk/pdbsum/2ye0 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=2ye0 ProSAT]</span></td></tr> | |||
</table> | |||
== Function == | |||
[https://www.uniprot.org/uniprot/GFP_AEQVI GFP_AEQVI] Energy-transfer acceptor. Its role is to transduce the blue chemiluminescence of the protein aequorin into green fluorescent light by energy transfer. Fluoresces in vivo upon receiving energy from the Ca(2+)-activated photoprotein aequorin. | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
Cyan variants of green fluorescent protein are widely used as donors in Forster resonance energy transfer experiments. The popular, but modestly bright, Enhanced Cyan Fluorescent Protein (ECFP) was sequentially improved into the brighter variants Super Cyan Fluorescent Protein 3A (SCFP3A) and mTurquoise, the latter exhibiting a high-fluorescence quantum yield and a long mono-exponential fluorescence lifetime. Here we combine X-ray crystallography and excited-state calculations to rationalize these stepwise improvements. The enhancement originates from stabilization of the seventh beta-strand and the strengthening of the sole chromophore-stabilizing hydrogen bond. The structural analysis highlighted one suboptimal internal residue, which was subjected to saturation mutagenesis combined with fluorescence lifetime-based screening. This resulted in mTurquoise2, a brighter variant with faster maturation, high photostability, longer mono-exponential lifetime and the highest quantum yield measured for a monomeric fluorescent protein. Together, these properties make mTurquoise2 the preferable cyan variant of green fluorescent protein for long-term imaging and as donor for Forster resonance energy transfer to a yellow fluorescent protein. | |||
Structure-guided evolution of cyan fluorescent proteins towards a quantum yield of 93%.,Goedhart J, von Stetten D, Noirclerc-Savoye M, Lelimousin M, Joosen L, Hink MA, van Weeren L, Gadella TW Jr, Royant A Nat Commun. 2012 Mar 20;3:751. doi: 10.1038/ncomms1738. PMID:22434194<ref>PMID:22434194</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
</div> | |||
<div class="pdbe-citations 2ye0" style="background-color:#fffaf0;"></div> | |||
==See Also== | |||
*[[Green Fluorescent Protein 3D structures|Green Fluorescent Protein 3D structures]] | |||
== References == | |||
<references/> | |||
__TOC__ | |||
</StructureSection> | |||
[[Category: Aequorea victoria]] | |||
[[Category: Large Structures]] | |||
[[Category: Gadella TWJ]] | |||
[[Category: Goedhart J]] | |||
[[Category: Hink MA]] | |||
[[Category: Joosen L]] | |||
[[Category: Lelimousin M]] | |||
[[Category: Noirclerc-Savoye M]] | |||
[[Category: Royant A]] | |||
[[Category: Van Weeren L]] | |||
[[Category: Von Stetten D]] | |||
Latest revision as of 11:13, 23 August 2023
X-ray structure of the cyan fluorescent protein mTurquoise (K206A mutant)X-ray structure of the cyan fluorescent protein mTurquoise (K206A mutant)
Structural highlights
FunctionGFP_AEQVI Energy-transfer acceptor. Its role is to transduce the blue chemiluminescence of the protein aequorin into green fluorescent light by energy transfer. Fluoresces in vivo upon receiving energy from the Ca(2+)-activated photoprotein aequorin. Publication Abstract from PubMedCyan variants of green fluorescent protein are widely used as donors in Forster resonance energy transfer experiments. The popular, but modestly bright, Enhanced Cyan Fluorescent Protein (ECFP) was sequentially improved into the brighter variants Super Cyan Fluorescent Protein 3A (SCFP3A) and mTurquoise, the latter exhibiting a high-fluorescence quantum yield and a long mono-exponential fluorescence lifetime. Here we combine X-ray crystallography and excited-state calculations to rationalize these stepwise improvements. The enhancement originates from stabilization of the seventh beta-strand and the strengthening of the sole chromophore-stabilizing hydrogen bond. The structural analysis highlighted one suboptimal internal residue, which was subjected to saturation mutagenesis combined with fluorescence lifetime-based screening. This resulted in mTurquoise2, a brighter variant with faster maturation, high photostability, longer mono-exponential lifetime and the highest quantum yield measured for a monomeric fluorescent protein. Together, these properties make mTurquoise2 the preferable cyan variant of green fluorescent protein for long-term imaging and as donor for Forster resonance energy transfer to a yellow fluorescent protein. Structure-guided evolution of cyan fluorescent proteins towards a quantum yield of 93%.,Goedhart J, von Stetten D, Noirclerc-Savoye M, Lelimousin M, Joosen L, Hink MA, van Weeren L, Gadella TW Jr, Royant A Nat Commun. 2012 Mar 20;3:751. doi: 10.1038/ncomms1738. PMID:22434194[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. See AlsoReferences
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