3ST0

Engineered medium-affinity halide-binding protein derived from YFP: halide-free

3ST0 の概要

• エントリーDOI: 10.2210/pdb3st0/pdb
• 関連するPDBエントリー: 3SRY 3SS0 3SSH 3SSK 3SSL 3SSP 3SST 3SSV 3SSY 3SV5 3SVB 3SVC 3SVD 3SVE
• 分子名称: Green fluorescent protein, 1,2-ETHANEDIOL, FORMIC ACID, ... (4 entities in total)
• 機能のキーワード: beta barrel, luminescent protein, yellow fluorescent protein, imaging reagent, halide binding protein
• 由来する生物種: Aequorea victoria (Jellyfish)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 29415.82

構造登録者

Wang, W.,Grimley, J.S.,Beese, L.S.,Hellinga, H.W. (登録日: 2011-07-08, 公開日: 2012-07-11, 最終更新日: 2024-04-03)

主引用文献

Grimley, J.S.,Li, L.,Wang, W.,Wen, L.,Beese, L.S.,Hellinga, H.W.,Augustine, G.J.
Visualization of Synaptic Inhibition with an Optogenetic Sensor Developed by Cell-Free Protein Engineering Automation.
J.Neurosci., 33:16297-16309, 2013

PubMed Abstract: We describe an engineered fluorescent optogenetic sensor, SuperClomeleon, that robustly detects inhibitory synaptic activity in single, cultured mouse neurons by reporting intracellular chloride changes produced by exogenous GABA or inhibitory synaptic activity. Using a cell-free protein engineering automation methodology that bypasses gene cloning, we iteratively constructed, produced, and assayed hundreds of mutations in binding-site residues to identify improvements in Clomeleon, a first-generation, suboptimal sensor. Structural analysis revealed that these improvements involve halide contacts and distant side chain rearrangements. The development of optogenetic sensors that respond to neural activity enables cellular tracking of neural activity using optical, rather than electrophysiological, signals. Construction of such sensors using in vitro protein engineering establishes a powerful approach for developing new probes for brain imaging.

PubMed: 24107961
DOI: 10.1523/JNEUROSCI.4616-11.2013

実験手法

X-RAY DIFFRACTION (1.19 Å)