4TVY

Apo resorufin ligase

4TVY の概要

• エントリーDOI: 10.2210/pdb4tvy/pdb
• 関連するPDBエントリー: 3A7R 4TVW
• 分子名称: Lipoate-protein ligase A, 5-(3,7-dihydroxy-10H-phenoxazin-2-yl)pentanamide (3 entities in total)
• 機能のキーワード: e. coli lpla, computational enzyme design, transferase
• 由来する生物種: Escherichia coli
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 77335.08

構造登録者

Goldman, P.J.,Drennan, C.L. (登録日: 2014-06-28, 公開日: 2014-10-22, 最終更新日: 2023-09-27)

主引用文献

Liu, D.S.,Nivon, L.G.,Richter, F.,Goldman, P.J.,Deerinck, T.J.,Yao, J.Z.,Richardson, D.,Phipps, W.S.,Ye, A.Z.,Ellisman, M.H.,Drennan, C.L.,Baker, D.,Ting, A.Y.
Computational design of a red fluorophore ligase for site-specific protein labeling in living cells.
Proc.Natl.Acad.Sci.USA, 111:E4551-E4559, 2014

PubMed Abstract: Chemical fluorophores offer tremendous size and photophysical advantages over fluorescent proteins but are much more challenging to target to specific cellular proteins. Here, we used Rosetta-based computation to design a fluorophore ligase that accepts the red dye resorufin, starting from Escherichia coli lipoic acid ligase. X-ray crystallography showed that the design closely matched the experimental structure. Resorufin ligase catalyzed the site-specific and covalent attachment of resorufin to various cellular proteins genetically fused to a 13-aa recognition peptide in multiple mammalian cell lines and in primary cultured neurons. We used resorufin ligase to perform superresolution imaging of the intermediate filament protein vimentin by stimulated emission depletion and electron microscopies. This work illustrates the power of Rosetta for major redesign of enzyme specificity and introduces a tool for minimally invasive, highly specific imaging of cellular proteins by both conventional and superresolution microscopies.

PubMed: 25313043
DOI: 10.1073/pnas.1404736111

実験手法

X-RAY DIFFRACTION (2.151 Å)