6M63

Crystal structure of a cAMP sensor G-Flamp1.

Summary for 6M63

• Entry DOI: 10.2210/pdb6m63/pdb
• Descriptor: Chimera of Cyclic nucleotide-gated potassium channel mll3241 and Yellow fluorescent protein, ADENOSINE-3',5'-CYCLIC-MONOPHOSPHATE (3 entities in total)
• Functional Keywords: gfp, cnbd, complex, signaling protein
• Biological source: Mesorhizobium japonicum MAFF 303099; More
• Total number of polymer chains: 2
• Total formula weight: 85899.58

Authors

Zhou, Z.,Chen, S.,Wang, L.,Chu, J. (deposition date: 2020-03-12, release date: 2021-09-22, Last modification date: 2024-11-06)

Primary citation

Wang, L.,Wu, C.,Peng, W.,Zhou, Z.,Zeng, J.,Li, X.,Yang, Y.,Yu, S.,Zou, Y.,Huang, M.,Liu, C.,Chen, Y.,Li, Y.,Ti, P.,Liu, W.,Gao, Y.,Zheng, W.,Zhong, H.,Gao, S.,Lu, Z.,Ren, P.G.,Ng, H.L.,He, J.,Chen, S.,Xu, M.,Li, Y.,Chu, J.
A high-performance genetically encoded fluorescent indicator for in vivo cAMP imaging.
Nat Commun, 13:5363-5363, 2022

PubMed Abstract: cAMP is a key second messenger that regulates diverse cellular functions including neural plasticity. However, the spatiotemporal dynamics of intracellular cAMP in intact organisms are largely unknown due to low sensitivity and/or brightness of current genetically encoded fluorescent cAMP indicators. Here, we report the development of the new circularly permuted GFP (cpGFP)-based cAMP indicator G-Flamp1, which exhibits a large fluorescence increase (a maximum ΔF/F of 1100% in HEK293T cells), decent brightness, appropriate affinity (a K of 2.17 μM) and fast response kinetics (an association and dissociation half-time of 0.20 and 0.087 s, respectively). Furthermore, the crystal structure of the cAMP-bound G-Flamp1 reveals one linker connecting the cAMP-binding domain to cpGFP adopts a distorted β-strand conformation that may serve as a fluorescence modulation switch. We demonstrate that G-Flamp1 enables sensitive monitoring of endogenous cAMP signals in brain regions that are implicated in learning and motor control in living organisms such as fruit flies and mice.

PubMed: 36097007
DOI: 10.1038/s41467-022-32994-7

Experimental method

X-RAY DIFFRACTION (2.25 Å)