4EDO

Crystal structure of far-red fluorescent protein eqFP650

4EDO の概要

• エントリーDOI: 10.2210/pdb4edo/pdb
• 関連するPDBエントリー: 3BXB 3IP2 3PJ7 4EDS
• 分子名称: far-red fluorescent protein eqFP650 (2 entities in total)
• 機能のキーワード: beta barrel, fluorescent protein
• 由来する生物種: Entacmaea quadricolor
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 54553.88

構造登録者

Pletnev, S.,Pletnev, V.Z.,Pletneva, N.V. (登録日: 2012-03-27, 公開日: 2012-09-05, 最終更新日: 2025-03-26)

主引用文献

Pletnev, S.,Pletneva, N.V.,Souslova, E.A.,Chudakov, D.M.,Lukyanov, S.,Wlodawer, A.,Dauter, Z.,Pletnev, V.
Structural basis for bathochromic shift of fluorescence in far-red fluorescent proteins eqFP650 and eqFP670.
Acta Crystallogr.,Sect.D, 68:1088-1097, 2012

PubMed Abstract: The crystal structures of the far-red fluorescent proteins (FPs) eqFP650 (λ(ex)(max)/λ(em)(max) 592/650 nm) and eqFP670 (λ(ex)(max)/λ(em)(max) 605/670 nm), the successors of the far-red FP Katushka (λ(ex)(max)/λ(em)(max) 588/635 nm), have been determined at 1.8 and 1.6 Å resolution, respectively. An examination of the structures demonstrated that there are two groups of changes responsible for the bathochromic shift of excitation/emission bands of these proteins relative to their predecessor. The first group of changes resulted in an increase of hydrophilicity at the acylimine site of the chromophore due to the presence of one and three water molecules in eqFP650 and eqFP670, respectively. These water molecules provide connection of the chromophore with the protein scaffold via hydrogen bonds causing an ~15 nm bathochromic shift of the eqFP650 and eqFP670 emission bands. The second group of changes observed in eqFP670 arises from substitution of both Ser143 and Ser158 by asparagines. Asn143 and Asn158 of eqFP670 are hydrogen bonded with each other, as well as with the protein scaffold and with the p-hydroxyphenyl group of the chromophore, resulting in an additional ~20 nm bathochromic shift of the eqFP670 emission band as compared to eqFP650. The role of the observed structural changes was verified by mutagenesis.

PubMed: 22948909
DOI: 10.1107/S0907444912020598

実験手法

X-RAY DIFFRACTION (1.8 Å)