2Y0G

X-ray structure of Enhanced Green Fluorescent Protein (EGFP)

2Y0G の概要

• エントリーDOI: 10.2210/pdb2y0g/pdb
• 関連するPDBエントリー: 1B9C 1BFP 1C4F 1CV7 1EMA 1EMB 1EMC 1EME 1EMF 1EMG 1EMK 1EML 1EMM 1F09 1F0B 1GFL 1H6R 1HCJ 1HUY 1JBY 1JBZ 1JC0 1JC1 1KP5 1KYP 1KYR 1KYS 1MYW 1Q4A 1Q4B 1Q4C 1Q4D 1Q4E 1Q73 1QXT 1QY3 1QYF 1QYO 1QYQ 1RM9 1RMM 1RMO 1RMP 1RRX 1W7S 1W7T 1W7U 1YFP 1YHG 1YHH 1YHI 1YJ2 1YJF 1Z1P 1Z1Q 2AH8 2AHA 2B3P 2B3Q 2EMD 2EMN 2EMO 2FWQ 2FZU 2WSN 2WSO 2WUR 2YFP
• 分子名称: GREEN FLUORESCENT PROTEIN (2 entities in total)
• 機能のキーワード: fluorescent 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: 28569.18

構造登録者

Royant, A.,Noirclerc-Savoye, M. (登録日: 2010-12-02, 公開日: 2011-04-27, 最終更新日: 2024-11-06)

主引用文献

Royant, A.,Noirclerc-Savoye, M.
Stabilizing Role of Glutamic Acid 222 in the Structure of Enhanced Green Fluorescent Protein.
J.Struct.Biol., 174:385-, 2011

PubMed Abstract: Enhanced Green Fluorescent Protein (EGFP) is a variant of wild-type Green Fluorescent Protein from the jellyfish Aequorea victoria, whose mutations S65T and F64L increase brightness and folding efficiency. EGFP is extensively used in cell biology and biochemistry as a colocalization or expression reporter. Surprisingly, the structure of this very popular protein has not been determined yet. We report here its crystallographic structure at 1.5Å resolution which shows significant differences in the vicinity of residue 64 and of the chromophore. In particular, two conformations are observed for the key residue glutamic acid 222, in apparent contradiction with the single fluorescence lifetime of the protein. We then show that X-ray induced decarboxylation of Glu222 during diffraction data collection results in the disruption of a hydrogen-bond network near the chromophore. Using single-crystal microspectrophotometry, we demonstrate that this correlates with a significant loss of the fluorescence properties. We thus propose a mechanism of bleaching of the protein at low temperature. Taken together, these two sets of results highlight the stabilizing role of Glu222 to the chromophore cavity of EGFP.

PubMed: 21335090
DOI: 10.1016/J.JSB.2011.02.004

実験手法

X-RAY DIFFRACTION (1.5 Å)