2vae
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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 Oct 25;. PMID:17962222
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