1H6R

The oxidized state of a redox sensitive variant of green fluorescent protein

Summary for 1H6R

• Entry DOI: 10.2210/pdb1h6r/pdb
• Related: 1B9C 1BFP 1C4F 1EMB 1EMC 1EME 1EMF 1EMG 1EMK 1EML 1EMM 1F09 1F0B 1HCJ 1YFP 2EMD 2EMN 2EMO
• Descriptor: GREEN FLUORESCENT PROTEIN, CHLORIDE ION (3 entities in total)
• Functional Keywords: luminescent protein, luminescence, green fluorescent protein, yellow-emission
• Biological source: AEQUOREA VICTORIA (JELLYFISH)
• Total number of polymer chains: 3
• Total formula weight: 80904.35

Authors

Ostergaard, H.,Henriksen, A.,Hansen, F.G.,Winther, J.R. (deposition date: 2001-06-22, release date: 2001-11-15, Last modification date: 2023-12-13)

Primary citation

Ostergaard, H.,Henriksen, A.,Hansen, F.G.,Winther, J.R.
Shedding Light on Disulfide Bond Formation: Engineering a Redox Switch in Green Fluorescent Protein
Embo J., 20:5853-, 2001

PubMed Abstract: To visualize the formation of disulfide bonds in living cells, a pair of redox-active cysteines was introduced into the yellow fluorescent variant of green fluorescent protein. Formation of a disulfide bond between the two cysteines was fully reversible and resulted in a >2-fold decrease in the intrinsic fluorescence. Inter conversion between the two redox states could thus be followed in vitro as well as in vivo by non-invasive fluorimetric measurements. The 1.5 A crystal structure of the oxidized protein revealed a disulfide bond-induced distortion of the beta-barrel, as well as a structural reorganization of residues in the immediate chromophore environment. By combining this information with spectroscopic data, we propose a detailed mechanism accounting for the observed redox state-dependent fluorescence. The redox potential of the cysteine couple was found to be within the physiological range for redox-active cysteines. In the cytoplasm of Escherichia coli, the protein was a sensitive probe for the redox changes that occur upon disruption of the thioredoxin reductive pathway.

PubMed: 11689426
DOI: 10.1093/EMBOJ/20.21.5853

Experimental method

X-RAY DIFFRACTION (1.5 Å)