6V9B

Co-crystal structure of the fluorogenic Mango-IV homodimer bound to TO1-Biotin

6V9B の概要

• エントリーDOI: 10.2210/pdb6v9b/pdb
• 分子名称: RNA (28-MER), ~{N}-methyl-2-[2-[(~{E})-(1-methylquinolin-4-ylidene)methyl]-1,3-benzothiazol-3-yl]ethanamide, POTASSIUM ION (3 entities in total)
• 機能のキーワード: rna, aptamer, fluorescence
• 由来する生物種: synthetic construct
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 19710.91

構造登録者

Trachman, R.J.,Ferre-D'Amare, A.R. (登録日: 2019-12-13, 公開日: 2020-05-20, 最終更新日: 2024-03-06)

主引用文献

Trachman 3rd, R.J.,Cojocaru, R.,Wu, D.,Piszczek, G.,Ryckelynck, M.,Unrau, P.J.,Ferre-D'Amare, A.R.
Structure-Guided Engineering of the Homodimeric Mango-IV Fluorescence Turn-on Aptamer Yields an RNA FRET Pair.
Structure, 28:776-785.e3, 2020

PubMed Abstract: Fluorescent RNA aptamers have been used in cells as biosensor reporters and tags for tracking transcripts. Recently, combined SELEX and microfluidic fluorescence sorting yielded three aptamers that activate fluorescence of TO1-Biotin: Mango-II, Mango-III, and Mango-IV. Of these, Mango-IV was best at imaging RNAs in both fixed and live mammalian cells. To understand how Mango-IV achieves activity in cells, we determined its crystal structure complexed with TO1-Biotin. The structure reveals a domain-swapped homodimer with two independent G-quadruplex fluorophore binding pockets. Structure-based analyses indicate that the Mango-IV core has relaxed fluorophore specificity, and a tendency to reorganize binding pocket residues. These molecular properties may endow it with robustness in the cellular milieu. Based on the domain-swapped structure, heterodimers between Mango-IV and the fluorescent aptamer iSpinach, joined by Watson-Crick base pairing, were constructed. These exhibited FRET between their respective aptamer-activated fluorophores, advancing fluorescent aptamer technology toward multi-color, RNA-based imaging of RNA coexpression and colocalization.

PubMed: 32386573
DOI: 10.1016/j.str.2020.04.007

実験手法

X-RAY DIFFRACTION (2.4 Å)