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Replacement of Highly Conserved E222 by the Photostable Non‐photoconvertible Histidine in GFP
Author(s) -
Auerbach Dagmar,
Klein Martin,
Franz Silke,
Carius Yvonne,
Lancaster C. Roy D.,
Jung Gregor
Publication year - 2014
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201402075
Subject(s) - green fluorescent protein , histidine , fluorescence , chemistry , biophysics , glutamic acid , amino acid , biochemistry , biology , gene , physics , quantum mechanics
The widely used green fluorescent protein (GFP) decarboxylates upon irradiation; this involves removal of the acidic function of the glutamic acid at position 222, thereby resulting in the irreversible photoconversion of GFP. To suppress this phenomenon, the photostable, non‐photoconvertible histidine was introduced at position 222 in GFP. The variant E222H shows negligible photodynamic processes and high expression yield. In addition, the stable and bright fluorescence over a wide pH range makes the E222H protein an alternative for GFP in fluorescence imaging and spectroscopy. Other fluorescent proteins are predicted to benefit from replacement of the catalytic glutamic acid by histidine.