3ST0

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

Summary for 3ST0

• Entry DOI: 10.2210/pdb3st0/pdb
• Related: 3SRY 3SS0 3SSH 3SSK 3SSL 3SSP 3SST 3SSV 3SSY 3SV5 3SVB 3SVC 3SVD 3SVE
• Descriptor: Green fluorescent protein, 1,2-ETHANEDIOL, FORMIC ACID, ... (4 entities in total)
• Functional Keywords: beta barrel, luminescent protein, yellow fluorescent protein, imaging reagent, halide binding protein
• Biological source: Aequorea victoria (Jellyfish)
• Total number of polymer chains: 1
• Total formula weight: 29415.82

Authors

Wang, W.,Grimley, J.S.,Beese, L.S.,Hellinga, H.W. (deposition date: 2011-07-08, release date: 2012-07-11, Last modification date: 2024-04-03)

Primary citation

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

Experimental method

X-RAY DIFFRACTION (1.19 Å)