2vae: Difference between revisions
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==Overview== | ==Overview== | ||
The red fluorescent protein DsRed has been extensively engineered for use | 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 Structure== | ==About this Structure== | ||
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==Reference== | ==Reference== | ||
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 Oct 25 | 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:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=17962222 17962222] | ||
[[Category: Discosoma sp.]] | [[Category: Discosoma sp.]] | ||
[[Category: Single protein]] | [[Category: Single protein]] | ||
[[Category: Bevis, B.]] | [[Category: Bevis, B.]] | ||
[[Category: Downing, M | [[Category: Downing, M E.]] | ||
[[Category: Glick, B | [[Category: Glick, B S.]] | ||
[[Category: Keenan, R | [[Category: Keenan, R J.]] | ||
[[Category: Khuong, N.]] | [[Category: Khuong, N.]] | ||
[[Category: Strack, R | [[Category: Strack, R L.]] | ||
[[Category: Strongin, D | [[Category: Strongin, D E.]] | ||
[[Category: Sundaram, K.]] | [[Category: Sundaram, K.]] | ||
[[Category: EDO]] | [[Category: EDO]] | ||
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[[Category: photoprotein]] | [[Category: photoprotein]] | ||
''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 18:54:35 2008'' | ||
Revision as of 19:54, 21 February 2008
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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. with as ligand. Known structural/functional Sites: , , , , , , , , , , , , , , , , , , , , , , , , , and . 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
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