4Q7U

Crystal structure of photoswitchable fluorescent protein PSmOrange2

4Q7U の概要

• エントリーDOI: 10.2210/pdb4q7u/pdb
• 関連するPDBエントリー: 2H5O 4Q7R 4Q7T
• 分子名称: PSmOrange2, GLYCEROL (3 entities in total)
• 機能のキーワード: beta-barrel, photoswitchable fluorescent protein, chromophore, fluorescent protein
• 由来する生物種: Discosoma sp. (Sea anemone)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 28244.95

構造登録者

Malashkevich, V.N.,Pletnev, S.,Almo, S.C. (登録日: 2014-04-25, 公開日: 2014-07-09, 最終更新日: 2023-12-06)

主引用文献

Pletnev, S.,Shcherbakova, D.M.,Subach, O.M.,Pletneva, N.V.,Malashkevich, V.N.,Almo, S.C.,Dauter, Z.,Verkhusha, V.V.
Orange Fluorescent Proteins: Structural Studies of LSSmOrange, PSmOrange and PSmOrange2.
Plos One, 9:e99136-e99136, 2014

PubMed Abstract: A structural analysis of the recently developed orange fluorescent proteins with novel phenotypes, LSSmOrange (λex/λem at 437/572 nm), PSmOrange (λex/λem at 548/565 nm and for photoconverted form at 636/662 nm) and PSmOrange2 (λex/λem at 546/561 nm and for photoconverted form at 619/651 nm), is presented. The obtained crystallographic structures provide an understanding of how the ensemble of a few key mutations enabled special properties of the orange FPs. While only a single Ile161Asp mutation, enabling excited state proton transfer, is critical for LSSmOrange, other substitutions provide refinement of its special properties and an exceptional 120 nm large Stokes shift. Similarly, a single Gln64Leu mutation was sufficient to cause structural changes resulting in photoswitchability of PSmOrange, and only one additional substitution (Phe65Ile), yielding PSmOrange2, was enough to greatly decrease the energy of photoconversion and increase its efficiency of photoswitching. Fluorescence of photoconverted PSmOrange and PSmOrange2 demonstrated an unexpected bathochromic shift relative to the fluorescence of classic red FPs, such as DsRed, eqFP578 and zFP574. The structural changes associated with this fluorescence shift are of considerable value for the design of advanced far-red FPs. For this reason the chromophore transformations accompanying photoconversion of the orange FPs are discussed.

PubMed: 24960050
DOI: 10.1371/journal.pone.0099136

実験手法

X-RAY DIFFRACTION (1.3 Å)