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Variants of DsRed fluorescent protein: Development of a copper sensor
Author(s) -
Eli Pharhad,
Chakrabartty Avijit
Publication year - 2006
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1110/ps.062239206
Subject(s) - copper , fluorescence , binding constant , mutant , metal , copper protein , quenching (fluorescence) , mutagenesis , chemistry , metal ions in aqueous solution , biophysics , photochemistry , crystallography , analytical chemistry (journal) , binding site , biochemistry , biology , gene , chromatography , physics , organic chemistry , quantum mechanics
The fluorescence quenching of drFP583 (DsRed) protein by metal ions was investigated. CuSO 4 reversibly and pH dependently quenched the red emission at 583 nm of drFP583. The copper binding constant was 15 mM. Following random mutagenesis, blue‐ and red‐shifted mutants of drFP583 were generated, and their metal sensing properties were examined. Mutant gRF possessed properties similar to green fluorescent protein and had a 18 mM copper binding constant. Mutant Rmu162 had an extraordinary red‐shifted emission peak at 620 nm. A third mutant, Rmu13, had dual emission peaks at 500 nm and 583 nm and possessed the properties of a copper sensor with a binding constant of 11 mM.