2vae: Difference between revisions

Revision as of 18:29, 4 May 2008

FAST MATURING RED FLUORESCENT PROTEIN, DSRED.T4

OverviewOverview

The red fluorescent protein DsRed has been extensively engineered for use as an in vivo research tool. In fast maturing DsRed variants, the chromophore maturation half-time is approximately 40 min, compared to approximately 12 h for wild-type DsRed. Further, DsRed has been converted from a tetramer into a monomer, a task that entailed mutating approximately 20% of the amino acids. These engineered variants of DsRed have proven extremely valuable for biomedical research, but the structural basis for the improved characteristics has not been thoroughly investigated. Here we present a 1.7 A crystal structure of the fast maturing tetrameric variant DsRed.T4. We also present a biochemical characterization and 1.6 A crystal structure of the monomeric variant DsRed.M1, also known as DsRed-Monomer. Analysis of the crystal structures suggests that rearrangements of Ser69 and Glu215 contribute to fast maturation, and that positioning of the Lys70 side chain modulates fluorescence quantum yield. Despite the 45 mutations in DsRed.M1 relative to wild-type DsRed, there is a root-mean-square deviation of only 0.3 A between the two structures. We propose that novel intramolecular interactions in DsRed.M1 partially compensate for the loss of intermolecular interactions found in the tetramer.

About this StructureAbout this Structure

2VAE is a Single protein structure of sequence from Discosoma sp.. Full crystallographic information is available from OCA.

ReferenceReference

Structural rearrangements near the chromophore influence the maturation speed and brightness of DsRed variants., Strongin DE, Bevis B, Khuong N, Downing ME, Strack RL, Sundaram K, Glick BS, Keenan RJ, Protein Eng Des Sel. 2007 Nov;20(11):525-34. Epub 2007 Oct 25. PMID:17962222 Page seeded by OCA on Sun May 4 18:29:50 2008

Proteopedia Page Contributors and Editors (what is this?)Proteopedia Page Contributors and Editors (what is this?)

OCA

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 [[Image:2vae.jpg|left|200px]]
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 '''FAST MATURING RED FLUORESCENT PROTEIN, DSRED.T4'''
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 [[Category: Strongin, D E.]]
 [[Category: Sundaram, K.]]
-[[Category: chromophore]]
+[[Category: Chromophore]]
-[[Category: dsred]]
+[[Category: Dsred]]
-[[Category: fast maturing]]
+[[Category: Fast maturing]]
-[[Category: fluorescent protein]]
+[[Category: Fluorescent protein]]
-[[Category: gfp-like]]
+[[Category: Gfp-like]]
-[[Category: luminescence]]
+[[Category: Luminescence]]
-[[Category: photoprotein]]
+[[Category: Photoprotein]]
+''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sun May  4 18:29:50 2008''
-''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Mon Mar 31 05:10:19 2008''