1huy: Difference between revisions

Revision as of 19:15, 2 May 2008

CRYSTAL STRUCTURE OF CITRINE, AN IMPROVED YELLOW VARIANT OF GREEN FLUORESCENT PROTEIN

OverviewOverview

Yellow mutants of the green fluorescent protein (YFP) are crucial constituents of genetically encoded indicators of signal transduction and fusions to monitor protein-protein interactions. However, previous YFPs show excessive pH sensitivity, chloride interference, poor photostability, or poor expression at 37 degrees C. Protein evolution in Escherichia coli has produced a new YFP named Citrine, in which the mutation Q69M confers a much lower pK(a) (5.7) than for previous YFPs, indifference to chloride, twice the photostability of previous YFPs, and much better expression at 37 degrees C and in organelles. The halide resistance is explained by a 2.2-A x-ray crystal structure of Citrine, showing that the methionine side chain fills what was once a large halide-binding cavity adjacent to the chromophore. Insertion of calmodulin within Citrine or fusion of cyan fluorescent protein, calmodulin, a calmodulin-binding peptide and Citrine has generated improved calcium indicators. These chimeras can be targeted to multiple cellular locations and have permitted the first single-cell imaging of free [Ca(2+)] in the Golgi. Citrine is superior to all previous YFPs except when pH or halide sensitivity is desired and is particularly advantageous within genetically encoded fluorescent indicators of physiological signals.

About this StructureAbout this Structure

1HUY is a Single protein structure of sequence from Aequorea victoria. Full crystallographic information is available from OCA.

ReferenceReference

Reducing the environmental sensitivity of yellow fluorescent protein. Mechanism and applications., Griesbeck O, Baird GS, Campbell RE, Zacharias DA, Tsien RY, J Biol Chem. 2001 Aug 3;276(31):29188-94. Epub 2001 May 31. PMID:11387331 Page seeded by OCA on Fri May 2 19:15:28 2008

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OCA

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 [[Image:1huy.jpg|left|200px]]
- {{Structure
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+The line below this paragraph, containing "STRUCTURE_1huy", creates the "Structure Box" on the page.
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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_1huy|  PDB=1huy  |  SCENE=  }}
-|RELATEDENTRY=[[1yfp|1YFP]], [[2yfp|2YFP]], [[1f09|1F09]], [[1f0b|1F0B]]
-|RESOURCES=<span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=1huy FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=1huy OCA], [http://www.ebi.ac.uk/pdbsum/1huy PDBsum], [http://www.rcsb.org/pdb/explore.do?structureId=1huy RCSB]</span>
-}}
 '''CRYSTAL STRUCTURE OF CITRINE, AN IMPROVED YELLOW VARIANT OF GREEN FLUORESCENT PROTEIN'''
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 [[Category: Tsien, R Y.]]
 [[Category: Zacharias, D A.]]
-[[Category: beta barrel]]
+[[Category: Beta barrel]]
-[[Category: chromophore]]
+[[Category: Chromophore]]
+''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Fri May  2 19:15:28 2008''
-''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sun Mar 30 21:10:46 2008''