5JZK

The Structure of Ultra Stable Green Fluorescent Protein

5JZK の概要

• エントリーDOI: 10.2210/pdb5jzk/pdb
• 関連するPDBエントリー: 5JZL
• 分子名称: Green fluorescent protein, NITRATE ION, 1,2-ETHANEDIOL, ... (6 entities in total)
• 機能のキーワード: green fluorescent protein, gfp, thermostable, fluorescent protein
• 由来する生物種: Aequorea victoria (Jellyfish)
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 56714.98

構造登録者

Yong, K.J.,Gunn, N.J.,Scott, D.J.,Griffin, M.D.W. (登録日: 2016-05-17, 公開日: 2017-12-06, 最終更新日: 2024-10-30)

主引用文献

Scott, D.J.,Gunn, N.J.,Yong, K.J.,Wimmer, V.C.,Veldhuis, N.A.,Challis, L.M.,Haidar, M.,Petrou, S.,Bathgate, R.A.D.,Griffin, M.D.W.
A Novel Ultra-Stable, Monomeric Green Fluorescent Protein For Direct Volumetric Imaging of Whole Organs Using CLARITY.
Sci Rep, 8:667-667, 2018

PubMed Abstract: Recent advances in thick tissue clearing are enabling high resolution, volumetric fluorescence imaging of complex cellular networks. Fluorescent proteins (FPs) such as GFP, however, can be inactivated by the denaturing chemicals used to remove lipids in some tissue clearing methods. Here, we solved the crystal structure of a recently engineered ultra-stable GFP (usGFP) and propose that the two stabilising mutations, Q69L and N164Y, act to improve hydrophobic packing in the core of the protein and facilitate hydrogen bonding networks at the surface, respectively. usGFP was found to dimerise strongly, which is not desirable for some applications. A point mutation at the dimer interface, F223D, generated monomeric usGFP (muGFP). Neurons in whole mouse brains were virally transduced with either EGFP or muGFP and subjected to Clear Lipid-exchanged Acrylamide-hybridized Rigid Imaging/Immunostaining/In situ hybridization-compatible Tissue-hYdrogel (CLARITY) clearing. muGFP fluorescence was retained after CLARITY whereas EGFP fluorescence was highly attenuated, thus demonstrating muGFP is a novel FP suitable for applications where high fluorescence stability and minimal self-association are required.

PubMed: 29330459
DOI: 10.1038/s41598-017-18045-y

実験手法

X-RAY DIFFRACTION (1.9 Å)