5OB3

iSpinach aptamer

5OB3 の概要

• エントリーDOI: 10.2210/pdb5ob3/pdb
• 分子名称: RNA aptamer (69-MER), 4-(3,5-difluoro-4-hydroxybenzyl)-1,2-dimethyl-1H-imidazol-5-ol, POTASSIUM ION, ... (5 entities in total)
• 機能のキーワード: aptamer, ispinach, dfhbi, rna
• 由来する生物種: synthetic construct
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 22908.01

構造登録者

Fernandez-Millan, P.,Autour, A.,Westhof, E.,Ryckelynck, M. (登録日: 2017-06-26, 公開日: 2017-10-04, 最終更新日: 2024-01-17)

主引用文献

Fernandez-Millan, P.,Autour, A.,Ennifar, E.,Westhof, E.,Ryckelynck, M.
Crystal structure and fluorescence properties of the iSpinach aptamer in complex with DFHBI.
RNA, 23:1788-1795, 2017

PubMed Abstract: Fluorogenic RNA aptamers are short nucleic acids able to specifically interact with small molecules and strongly enhance their fluorescence upon complex formation. Among the different systems recently introduced, Spinach, an aptamer forming a fluorescent complex with the 3,5-difluoro-4-hydroxybenzylidene imidazolinone (DFHBI), is one of the most promising. Using random mutagenesis and ultrahigh-throughput screening, we recently developed iSpinach, an improved version of the aptamer, endowed with an increased folding efficiency and thermal stability. iSpinach is a shorter version of Spinach, comprising five mutations for which the exact role has not yet been deciphered. In this work, we cocrystallized a reengineered version of iSpinach in complex with the DFHBI and solved the X-ray structure of the complex at 2 Å resolution. Only a few mutations were required to optimize iSpinach production and crystallization, underlying the good folding capacity of the molecule. The measured fluorescence half-lives in the crystal were 60% higher than in solution. Comparisons with structures previously reported for Spinach sheds some light on the possible function of the different beneficial mutations carried by iSpinach.

PubMed: 28939697
DOI: 10.1261/rna.063008.117

実験手法

X-RAY DIFFRACTION (2.004 Å)