4XTQ

Crystal structure of a mutant (C20S) of a near-infrared fluorescent protein BphP1-FP

4XTQ の概要

• エントリーDOI: 10.2210/pdb4xtq/pdb
• 分子名称: BphP1-FP/C20S, 3-[2-[(Z)-[5-[(Z)-[(3R,4R)-3-ethenyl-4-methyl-5-oxidanylidene-pyrrolidin-2-ylidene]methyl]-3-(3-hydroxy-3-oxopropyl)-4-methyl-pyrrol-2-ylidene]methyl]-5-[(Z)-(4-ethenyl-3-methyl-5-oxidanylidene-pyrrol-2-ylidene)methyl]-4-methyl-1H-pyrrol-3-yl]propanoic acid, CHLORIDE ION, ... (4 entities in total)
• 機能のキーワード: bacteriophytochrome, bphp, biliverdin, near-infrared fluorescent protein, fluorescent protein
• 由来する生物種: Rhodopseudomonas palustris
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 36666.81

構造登録者

Pletnev, S.,Malashkevich, V.N. (登録日: 2015-01-23, 公開日: 2015-12-09, 最終更新日: 2024-12-25)

主引用文献

Shcherbakova, D.M.,Baloban, M.,Pletnev, S.,Malashkevich, V.N.,Xiao, H.,Dauter, Z.,Verkhusha, V.V.
Molecular Basis of Spectral Diversity in Near-Infrared Phytochrome-Based Fluorescent Proteins.
Chem.Biol., 22:1540-1551, 2015

PubMed Abstract: Near-infrared fluorescent proteins (NIR FPs) engineered from bacterial phytochromes (BphPs) are the probes of choice for deep-tissue imaging. Detection of several processes requires spectrally distinct NIR FPs. We developed an NIR FP, BphP1-FP, which has the most blue-shifted spectra and the highest fluorescence quantum yield among BphP-derived FPs. We found that these properties result from the binding of the biliverdin chromophore to a cysteine residue in the GAF domain, unlike natural BphPs and other BphP-based FPs. To elucidate the molecular basis of the spectral shift, we applied biochemical, structural and mass spectrometry analyses and revealed the formation of unique chromophore species. Mutagenesis of NIR FPs of different origins indicated that the mechanism of the spectral shift is general and can be used to design multicolor NIR FPs from other BphPs. We applied pairs of spectrally distinct point cysteine mutants to multicolor cell labeling and demonstrated that they perform well in model deep-tissue imaging.

PubMed: 26590639
DOI: 10.1016/j.chembiol.2015.10.007

実験手法

X-RAY DIFFRACTION (1.64 Å)