1xss
From Proteopedia
(New page: 200px<br /><applet load="1xss" size="450" color="white" frame="true" align="right" spinBox="true" caption="1xss, resolution 1.6Å" /> '''Semi-rational enginee...) |
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| - | [[Image:1xss.gif|left|200px]]<br /><applet load="1xss" size=" | + | [[Image:1xss.gif|left|200px]]<br /><applet load="1xss" size="350" color="white" frame="true" align="right" spinBox="true" |
caption="1xss, resolution 1.6Å" /> | caption="1xss, resolution 1.6Å" /> | ||
'''Semi-rational engineering of a green-emitting coral fluorescent protein into an efficient highlighter.'''<br /> | '''Semi-rational engineering of a green-emitting coral fluorescent protein into an efficient highlighter.'''<br /> | ||
==Overview== | ==Overview== | ||
| - | Kaede is a natural photoconvertible fluorescent protein found in the coral | + | Kaede is a natural photoconvertible fluorescent protein found in the coral Trachyphyllia geoffroyi. It contains a tripeptide, His 62-Tyr 63-Gly 64, which acts as a green chromophore that is photoconvertible to red following (ultra-) violet irradiation. Here, we report the molecular cloning and crystal structure determination of a new fluorescent protein, KikG, from the coral Favia favus, and its in vitro evolution conferring green-to-red photoconvertibility. Substitution of the His 62-Tyr 63-Gly 64 sequence into the native protein provided only negligible photoconversion. On the basis of the crystal structure, semi-rational mutagenesis of the amino acids surrounding the chromophore was performed, leading to the generation of an efficient highlighter, KikGR. Within mammalian cells, KikGR is more efficiently photoconverted and is several-fold brighter in both the green and red states than Kaede. In addition, KikGR was successfully photoconverted using two-photon excitation microscopy at 760 nm, ensuring optical cell labelling with better spatial discrimination in thick and highly scattering tissues. |
==About this Structure== | ==About this Structure== | ||
| - | 1XSS is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/Favia_favus Favia favus] with MG and NA as [http://en.wikipedia.org/wiki/ligands ligands]. Full crystallographic information is available from [http:// | + | 1XSS is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/Favia_favus Favia favus] with <scene name='pdbligand=MG:'>MG</scene> and <scene name='pdbligand=NA:'>NA</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1XSS OCA]. |
==Reference== | ==Reference== | ||
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[[Category: fluorescent protein]] | [[Category: fluorescent protein]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 15:58:18 2008'' |
Revision as of 13:58, 21 February 2008
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Semi-rational engineering of a green-emitting coral fluorescent protein into an efficient highlighter.
Overview
Kaede is a natural photoconvertible fluorescent protein found in the coral Trachyphyllia geoffroyi. It contains a tripeptide, His 62-Tyr 63-Gly 64, which acts as a green chromophore that is photoconvertible to red following (ultra-) violet irradiation. Here, we report the molecular cloning and crystal structure determination of a new fluorescent protein, KikG, from the coral Favia favus, and its in vitro evolution conferring green-to-red photoconvertibility. Substitution of the His 62-Tyr 63-Gly 64 sequence into the native protein provided only negligible photoconversion. On the basis of the crystal structure, semi-rational mutagenesis of the amino acids surrounding the chromophore was performed, leading to the generation of an efficient highlighter, KikGR. Within mammalian cells, KikGR is more efficiently photoconverted and is several-fold brighter in both the green and red states than Kaede. In addition, KikGR was successfully photoconverted using two-photon excitation microscopy at 760 nm, ensuring optical cell labelling with better spatial discrimination in thick and highly scattering tissues.
About this Structure
1XSS is a Protein complex structure of sequences from Favia favus with and as ligands. Full crystallographic information is available from OCA.
Reference
Semi-rational engineering of a coral fluorescent protein into an efficient highlighter., Tsutsui H, Karasawa S, Shimizu H, Nukina N, Miyawaki A, EMBO Rep. 2005 Mar;6(3):233-8. PMID:15731765
Page seeded by OCA on Thu Feb 21 15:58:18 2008
Categories: Favia favus | Protein complex | Karasawa, S. | Miyawaki, A. | Nukina, N. | Shimizu, H. | Tsutsui, H. | MG | NA | Fluorescent protein
