8BGL

Structure of the dimeric rsCherryRev1.4

Summary for 8BGL

• Entry DOI: 10.2210/pdb8bgl/pdb
• Related: 8B65 8B7G
• Descriptor: PAmCherry1 protein, DI(HYDROXYETHYL)ETHER, PENTAETHYLENE GLYCOL, ... (5 entities in total)
• Functional Keywords: dimerization, reversibly switchable, red fluorescent proteins, disulfide bond, fluorescent protein
• Biological source: Discosoma sp.
• Total number of polymer chains: 2
• Total formula weight: 61595.45

Authors

Bui, T.Y.H.,Van Meervelt, L. (deposition date: 2022-10-28, release date: 2023-02-15, Last modification date: 2024-10-16)

Primary citation

Bui, T.Y.H.,Dedecker, P.,Van Meervelt, L.
An unusual disulfide-linked dimerization in the fluorescent protein rsCherryRev1.4.
Acta Crystallogr.,Sect.F, 79:38-44, 2023

PubMed Abstract: rsCherryRev1.4 has been reported as one of the reversibly photoswitchable variants of mCherry, and is an improved version with a faster off-switching speed and lower switching fatigue at high light intensities than its precursor rsCherryRev. However, rsCherryRev1.4 still has some limitations such as a tendency to dimerize as well as complex photophysical properties. Here, the crystal structure of rsCherryRev1.4 was determined at a resolution of 2 Å and it was discovered that it forms a dimer that shows disulfide bonding between the protomers. Mutagenesis, gel electrophoresis and size-exclusion chromatography strongly implicate Cys24 in this process. Replacing Cys24 in rsCherryRev1.4 resulted in a much lower tendency towards dimerization, while introducing Cys24 into mCherry correspondingly increased its dimerization. In principle, this finding opens the possibility of developing redox sensors based on controlled dimerization via disulfide cross-linking in fluorescent proteins, even though the actual application of engineering such sensors still requires additional research.

PubMed: 36748340
DOI: 10.1107/S2053230X23000572

Experimental method

X-RAY DIFFRACTION (2 Å)