3AKO

Crystal Structure of the Reassembled Venus

3AKO の概要

• エントリーDOI: 10.2210/pdb3ako/pdb
• 分子名称: Venus, SULFATE ION, 3,6,9,12,15,18,21-HEPTAOXATRICOSANE-1,23-DIOL, ... (5 entities in total)
• 機能のキーワード: venus, fluorescent protein, gfp
• 由来する生物種: Plant transformation vector pSITEII-4C1; 詳細
• タンパク質・核酸の鎖数: 8
• 化学式量合計: 122655.78

構造登録者

Isogai, M.,Tada, T. (登録日: 2010-07-15, 公開日: 2011-08-03, 最終更新日: 2026-03-18)

主引用文献

Isogai, M.,Kawamoto, Y.,Inahata, K.,Fukada, H.,Sugimoto, K.,Tada, T.
Structure and characteristics of reassembled fluorescent protein, a new insight into the reassembly mechanisms
Bioorg.Med.Chem.Lett., 21:3021-3024, 2011

PubMed Abstract: Bimolecular fluorescence complementation (BiFC) assay has been used widely to visualize protein-protein interactions in cells. However, there is a problem that fluorescent protein fragments have an ability to associate with each other independent of an interaction between proteins fused to the fragments. To facilitate the BiFC assay, we have attempted to determine the structure and characteristics of reassembled fluorescent protein, Venus. The anion-exchange chromatography showed an oligomer and a monomer of reassembled Venus. Our results suggested that the oligomer was formed by β-strands swapping without any serious steric clashes and was converted to the monomer. Crystal structure of reassembled Venus had an 11-stranded β-barrel fold, typical of GFP-derived fluorescent proteins. Based on the structural features, we have mutated to β-strand 7 and measured T(m) values. The results have revealed that the mutation influences the thermal stability of reassembled fluorescent complex.

PubMed: 21463942
DOI: 10.1016/j.bmcl.2011.03.039

実験手法

X-RAY DIFFRACTION (2.1 Å)