1HUY

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

1HUY の概要

• エントリーDOI: 10.2210/pdb1huy/pdb
• 関連するPDBエントリー: 1F09 1F0B 1YFP 2YFP
• 分子名称: GREEN FLUORESCENT PROTEIN (2 entities in total)
• 機能のキーワード: beta barrel, chromophore, luminescent protein
• 由来する生物種: Aequorea victoria
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 27319.88

構造登録者

Griesbeck, O.,Baird, G.S.,Campbell, R.E.,Zacharias, D.A.,Tsien, R.Y. (登録日: 2001-01-04, 公開日: 2001-07-04, 最終更新日: 2024-11-20)

主引用文献

Griesbeck, O.,Baird, G.S.,Campbell, R.E.,Zacharias, D.A.,Tsien, R.Y.
Reducing the environmental sensitivity of yellow fluorescent protein. Mechanism and applications
J.Biol.Chem., 276:29188-29194, 2001

PubMed Abstract: 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.

PubMed: 11387331
DOI: 10.1074/jbc.M102815200

実験手法

X-RAY DIFFRACTION (2.2 Å)