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1xss

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{{STRUCTURE_1xss| PDB=1xss | SCENE= }}
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'''Semi-rational engineering of a green-emitting coral fluorescent protein into an efficient highlighter.'''
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===Semi-rational engineering of a green-emitting coral fluorescent protein into an efficient highlighter.===
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==Overview==
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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.
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(as it appears on PubMed at http://www.pubmed.gov), where 15731765 is the PubMed ID number.
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{{ABSTRACT_PUBMED_15731765}}
==About this Structure==
==About this Structure==
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[[Category: Tsutsui, H.]]
[[Category: Tsutsui, H.]]
[[Category: Fluorescent protein]]
[[Category: Fluorescent protein]]
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sat May 3 15:28:02 2008''
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Mon Jul 28 00:55:56 2008''

Revision as of 21:55, 27 July 2008

Image:1xss.png

Template:STRUCTURE 1xss

Semi-rational engineering of a green-emitting coral fluorescent protein into an efficient highlighter.

Template:ABSTRACT PUBMED 15731765

About this Structure

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 Mon Jul 28 00:55:56 2008

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