2VAD

Monomeric red fluorescent protein, DsRed.M1

2VAD の概要

• エントリーDOI: 10.2210/pdb2vad/pdb
• 関連するPDBエントリー: 2VAE
• 分子名称: RED FLUORESCENT PROTEIN, ZINC ION, CHLORIDE ION, ... (5 entities in total)
• 機能のキーワード: dsred, gfp-like, monomeric, chromophore, luminescence, photoprotein, fluorescent protein
• 由来する生物種: DISCOSOMA SP. (SEA ANEMONE)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 25867.07

構造登録者

Strongin, D.E.,Bevis, B.,Khuong, N.,Downing, M.E.,Strack, R.L.,Sundaram, K.,Glick, B.S.,Keenan, R.J. (登録日: 2007-08-30, 公開日: 2007-11-06, 最終更新日: 2024-10-23)

主引用文献

Strongin, D.E.,Bevis, B.,Khuong, N.,Downing, M.E.,Strack, R.L.,Sundaram, K.,Glick, B.S.,Keenan, R.J.
Structural Rearrangements Near the Chromophore Influence the Maturation Speed and Brightness of Dsred Variants.
Protein Eng.Des.Sel., 20:525-, 2007

PubMed Abstract: 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.

PubMed: 17962222
DOI: 10.1093/PROTEIN/GZM046

実験手法

X-RAY DIFFRACTION (1.59 Å)