2y0g

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X-ray structure of Enhanced Green Fluorescent Protein (EGFP)X-ray structure of Enhanced Green Fluorescent Protein (EGFP)

Structural highlights

2y0g is a 1 chain structure with sequence from Aequorea victoria. The June 2014 RCSB PDB Molecule of the Month feature on GFP-like Proteins by David Goodsell is 10.2210/rcsb_pdb/mom_2014_6. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance.
Method:X-ray diffraction, Resolution 1.5Å
Ligands:
Resources:FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT

Function

GFP_AEQVI Energy-transfer acceptor. Its role is to transduce the blue chemiluminescence of the protein aequorin into green fluorescent light by energy transfer. Fluoresces in vivo upon receiving energy from the Ca(2+)-activated photoprotein aequorin.

Publication Abstract from PubMed

Enhanced Green Fluorescent Protein (EGFP) is a variant of wild-type Green Fluorescent Protein from the jellyfish Aequorea victoria, whose mutations S65T and F64L increase brightness and folding efficiency. EGFP is extensively used in cell biology and biochemistry as a colocalization or expression reporter. Surprisingly, the structure of this very popular protein has not been determined yet. We report here its crystallographic structure at 1.5A resolution which shows significant differences in the vicinity of residue 64 and of the chromophore. In particular, two conformations are observed for the key residue glutamic acid 222, in apparent contradiction with the single fluorescence lifetime of the protein. We then show that X-ray induced decarboxylation of Glu222 during diffraction data collection results in the disruption of a hydrogen-bond network near the chromophore. Using single-crystal microspectrophotometry, we demonstrate that this correlates with a significant loss of the fluorescence properties. We thus propose a mechanism of bleaching of the protein at low temperature. Taken together, these two sets of results highlight the stabilizing role of Glu222 to the chromophore cavity of EGFP.

Stabilizing role of glutamic acid 222 in the structure of Enhanced Green Fluorescent Protein.,Royant A, Noirclerc-Savoye M J Struct Biol. 2011 May;174(2):385-90. Epub 2011 Feb 16. PMID:21335090[1]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

See Also

References

  1. Royant A, Noirclerc-Savoye M. Stabilizing role of glutamic acid 222 in the structure of Enhanced Green Fluorescent Protein. J Struct Biol. 2011 May;174(2):385-90. Epub 2011 Feb 16. PMID:21335090 doi:10.1016/j.jsb.2011.02.004

2y0g, resolution 1.50Å

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