6LNP

Crystal structure of citrate Biosensor

Summary for 6LNP

• Entry DOI: 10.2210/pdb6lnp/pdb
• Descriptor: Fusion protein of Green fluorescent protein and Sensor histidine kinase CitA, CITRIC ACID (2 entities in total)
• Functional Keywords: biosensor, genetically encoded, citrate, fluorescent protein
• Biological source: Aequorea victoria; More
• Total number of polymer chains: 2
• Total formula weight: 90127.42

Authors

Wen, Y.,Campbell, R. (deposition date: 2019-12-31, release date: 2020-05-06, Last modification date: 2024-10-09)

Primary citation

Zhao, Y.,Shen, Y.,Wen, Y.,Campbell, R.E.
High-Performance Intensiometric Direct- and Inverse-Response Genetically Encoded Biosensors for Citrate.
Acs Cent.Sci., 6:1441-1450, 2020

PubMed Abstract: Motivated by the growing recognition of citrate as a central metabolite in a variety of biological processes associated with healthy and diseased cellular states, we have developed a series of high-performance genetically encoded citrate biosensors suitable for imaging of citrate concentrations in mammalian cells. The design of these biosensors was guided by structural studies of the citrate-responsive sensor histidine kinase and took advantage of the same conformational changes proposed to propagate from the binding domain to the catalytic domain. Following extensive engineering based on a combination of structure guided mutagenesis and directed evolution, we produced an inverse-response biosensor (Δ/ ≈ 18) designated Citroff1 and a direct-response biosensor (Δ/ ≈ 9) designated Citron1. We report the X-ray crystal structure of Citron1 and demonstrate the utility of both biosensors for qualitative and quantitative imaging of steady-state and pharmacologically perturbed citrate concentrations in live cells.

PubMed: 32875085
DOI: 10.1021/acscentsci.0c00518

Experimental method

X-RAY DIFFRACTION (2.993 Å)