1h6r
From Proteopedia
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THE OXIDIZED STATE OF A REDOX SENSITIVE VARIANT OF GREEN FLUORESCENT PROTEIN
Overview
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.
About this Structure
1H6R is a Single protein structure of sequence from Aequorea victoria with CL as ligand. Full crystallographic information is available from OCA.
Reference
Shedding light on disulfide bond formation: engineering a redox switch in green fluorescent protein., Ostergaard H, Henriksen A, Hansen FG, Winther JR, EMBO J. 2001 Nov 1;20(21):5853-62. PMID:11689426
Page seeded by OCA on Tue Nov 20 16:26:24 2007
