2yfp: Difference between revisions

Revision as of 19:11, 4 May 2008

STRUCTURE OF YELLOW-EMISSION VARIANT OF GFP

OverviewOverview

BACKGROUND: Because of its ability to spontaneously generate its own fluorophore, the green fluorescent protein (GFP) from the jellyfish Aequorea victoria is used extensively as a fluorescent marker in molecular and cell biology. The yellow fluorescent proteins (YFPs) have the longest wavelength emissions of all GFP variants examined to date. This shift in the spectrum is the result of a T203Y substitution (single-letter amino acid code), a mutation rationally designed on the basis of the X-ray structure of GFP S65T. RESULTS: We have determined the crystal structures of YFP T203Y/S65G/V68L/S72A and YFP H148G to 2.5 and 2.6 A resolution, respectively. Both structures show clear electron density for nearly coplanar pi-pi stacking between Tyr203 and the chromophore. The chromophore has been displaced by nearly 1 A in comparison to other available structures. Although the H148G mutation results in the generation of a solvent channel to the chromophore cavity, intense fluorescence is maintained. The chromophore in the intact protein can be titrated, and the two variants have pKa values of 7.0 (YFP) and 8.0 (YFP H148G). CONCLUSIONS: The observed red shift of the T203Y YFP variant is proposed to be mainly due to the additional polarizability of the pi-stacked Tyr203. The altered location of the chromophore suggests that the exact positions of nearby residues are not crucial for the chemistry of chromophore formation. The YFPs significantly extend the pH range over which GFPs may be employed as pH indicators in live cells.

About this StructureAbout this Structure

2YFP is a Single protein structure of sequence from Aequorea victoria. Full crystallographic information is available from OCA.

ReferenceReference

Structural basis of spectral shifts in the yellow-emission variants of green fluorescent protein., Wachter RM, Elsliger MA, Kallio K, Hanson GT, Remington SJ, Structure. 1998 Oct 15;6(10):1267-77. PMID:9782051 Page seeded by OCA on Sun May 4 19:11:33 2008

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OCA

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 [[Image:2yfp.jpg|left|200px]]
- {{Structure
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+The line below this paragraph, containing "STRUCTURE_2yfp", creates the "Structure Box" on the page.
-|SITE=
+You may change the PDB parameter (which sets the PDB file loaded into the applet)
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-|GENE= GFP ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=6100 Aequorea victoria])
+-->
-|DOMAIN=
+{{STRUCTURE_2yfp|  PDB=2yfp  |  SCENE=  }}
-|RELATEDENTRY=
-|RESOURCES=<span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=2yfp FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=2yfp OCA], [http://www.ebi.ac.uk/pdbsum/2yfp PDBsum], [http://www.rcsb.org/pdb/explore.do?structureId=2yfp RCSB]</span>
-}}
 '''STRUCTURE OF YELLOW-EMISSION VARIANT OF GFP'''
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 [[Category: Remington, S J.]]
 [[Category: Wachter, R M.]]
-[[Category: bioluminescence]]
+[[Category: Bioluminescence]]
-[[Category: fluorescent tag]]
+[[Category: Fluorescent tag]]
-[[Category: green fluorescent protein]]
+[[Category: Green fluorescent protein]]
-[[Category: photoactive protein]]
+[[Category: Photoactive protein]]
-[[Category: yellow-emission variant]]
+[[Category: Yellow-emission variant]]
+''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sun May  4 19:11:33 2008''
-''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Mon Mar 31 05:14:20 2008''