7ZBB

HaloTag with TRaQ-G-ctrl ligand

Summary for 7ZBB

• Entry DOI: 10.2210/pdb7zbb/pdb
• Descriptor: Haloalkane dehalogenase, (E)-[7-azanyl-10-[2-carboxy-5-[2-[2-(6-chloranylhexoxy)ethoxy]ethylcarbamoyl]phenyl]-5,5-dimethyl-benzo[b][1]benzosilin-3-ylidene]-methyl-azanium, GLYCEROL, ... (5 entities in total)
• Functional Keywords: halotag, haloalkane dehalogenase, silicon rhodamine, hydrolase
• Biological source: Rhodococcus sp.
• Total number of polymer chains: 2
• Total formula weight: 70568.33

Authors

Emmert, S.,Rivera-Fuentes, P.,Pojer, F.,Lau, K. (deposition date: 2022-03-23, release date: 2023-02-01, Last modification date: 2024-11-20)

Primary citation

Emmert, S.,Quargnali, G.,Thallmair, S.,Rivera-Fuentes, P.
A locally activatable sensor for robust quantification of organellar glutathione.
Nat.Chem., 15:1415-1421, 2023

PubMed Abstract: Glutathione (GSH) is the main determinant of intracellular redox potential and participates in multiple cellular signalling pathways. Achieving a detailed understanding of intracellular GSH homeostasis depends on the development of tools to map GSH compartmentalization and intra-organelle fluctuations. Here we present a GSH-sensing platform for live-cell imaging, termed targetable ratiometric quantitative GSH (TRaQ-G). This chemogenetic sensor possesses a unique reactivity turn-on mechanism, ensuring that the small molecule is only sensitive to GSH in a desired location. Furthermore, TRaQ-G can be fused to a fluorescent protein to give a ratiometric response. Using TRaQ-G fused to a redox-insensitive fluorescent protein, we demonstrate that the nuclear and cytosolic GSH pools are independently regulated during cell proliferation. This sensor was used in combination with a redox-sensitive fluorescent protein to quantify redox potential and GSH concentration simultaneously in the endoplasmic reticulum. Finally, by exchanging the fluorescent protein, we created a near-infrared, targetable and quantitative GSH sensor.

PubMed: 37322101
DOI: 10.1038/s41557-023-01249-3

Experimental method

X-RAY DIFFRACTION (1.95 Å)