6J7U

Crystal structure of blue fluorescent protein from metagenomic library in complex with NADPH

6J7U の概要

• エントリーDOI: 10.2210/pdb6j7u/pdb
• 分子名称: Blue fluorescent protein, NADPH DIHYDRO-NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE (3 entities in total)
• 機能のキーワード: dehydrogenase, bfp, oxidoreductase
• 由来する生物種: uncultured bacterium
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 110511.25

構造登録者

Seo, P.W.,Kim, J.S. (登録日: 2019-01-18, 公開日: 2019-09-18, 最終更新日: 2023-11-22)

主引用文献

Seo, P.W.,Jo, E.S.,You, S.H.,Cheong, D.E.,Kim, G.J.,Kim, J.S.
Structure-Guided Generation of a Redox-Independent Blue Fluorescent Protein from mBFP.
J.Mol.Biol., 431:3191-3202, 2019

PubMed Abstract: Fluorescent proteins, such as the green fluorescent protein, are used for detection of cellular components and events. However, green fluorescent protein and its derivatives have limited usage under anaerobic conditions and require a long maturation time. On the other hand, the NADPH-dependent blue fluorescent protein (BFP) without oxidative modification of residues is instantly functional in both aerobic and anaerobic systems. BFP proteins belong to a short-chain dehydrogenase/reductase (SDR) protein family, and their fluorescent property changes with reaction time in the presence of a substrate. With the aim of developing a better fluorescent reporter independent of redox state, we elucidated the crystal structure of a tetrameric mBFP from soil metagenomes with and without NADPH. Apart from the previously known regions, structure-guided mutational studies have identified several residues that contribute to the fluorescence of mBFP, including two aromatic residues (F97 and Y157) near the nicotinamide moiety of the bound NADPH. A single histidine mutation at Y157 (Y157H) has conferred more stabilized, time-independent fluorescence even in the presence of substrates. Furthermore, we discovered another SDR protein that can also emit blue fluorescence. These results open a new possibility for the development of BFP as a stable cellular reporter for widespread use, independent of subcellular environments.

PubMed: 31202883
DOI: 10.1016/j.jmb.2019.06.005

実験手法

X-RAY DIFFRACTION (2.302 Å)