8PM1

Structure of Chloroflexus aggregans flavin based fluorescent protein (CagFbFP) variant I52V A85Q

8PM1 の概要

• エントリーDOI: 10.2210/pdb8pm1/pdb
• 関連するPDBエントリー: 8PKY
• 分子名称: histidine kinase, FLAVIN MONONUCLEOTIDE (3 entities in total)
• 機能のキーワード: lov domain, fluorescent protein
• 由来する生物種: Chloroflexus aggregans
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 25800.50

構造登録者

Nikolaev, A.,Remeeva, A.,Gushchin, I. (登録日: 2023-06-27, 公開日: 2023-07-05, 最終更新日: 2024-01-17)

主引用文献

Nikolaev, A.,Tropina, E.V.,Boldyrev, K.N.,Maksimov, E.G.,Borshchevskiy, V.,Mishin, A.,Yudenko, A.,Kuzmin, A.,Kuznetsova, E.,Semenov, O.,Remeeva, A.,Gushchin, I.
Two distinct mechanisms of flavoprotein spectral tuning revealed by low-temperature and time-dependent spectroscopy.
Protein Sci., 33:e4851-e4851, 2024

PubMed Abstract: Flavins such as flavin mononucleotide or flavin adenine dinucleotide are bound by diverse proteins, yet have very similar spectra when in the oxidized state. Recently, we developed new variants of flavin-binding protein CagFbFP exhibiting notable blue (Q148V) or red (I52V A85Q) shifts of fluorescence emission maxima. Here, we use time-resolved and low-temperature spectroscopy to show that whereas the chromophore environment is static in Q148V, an additional protein-flavin hydrogen bond is formed upon photoexcitation in the I52V A85Q variant. Consequently, in Q148V, excitation, emission, and phosphorescence spectra are shifted, whereas in I52V A85Q, excitation and low-temperature phosphorescence spectra are relatively unchanged, while emission spectrum is altered. We also determine the x-ray structures of the two variants to reveal the flavin environment and complement the spectroscopy data. Our findings illustrate two distinct color-tuning mechanisms of flavin-binding proteins and could be helpful for the engineering of new variants with improved optical properties.

PubMed: 38038877
DOI: 10.1002/pro.4851

実験手法

X-RAY DIFFRACTION (1.5 Å)