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Tryptophan and tyrosine are the two that have obvious PET interaction with a fluorophore in protein or aqueous solution [1-6].Herein,we selected two commonly used fluorophores,Atto655 and Eosin-Y (Scheme 1),and studied their respective ultrafast fluorescence quenching dynamics in the presence of N-Acetyl-Tyrosine (AcTyr) and N-Acetyl-Tryptophan (AcTrp) in aqueous solution with femtosecond transient absorption spectroscopy [5,6].First,we measured the first electronic excited-state decays of Atto655 in different concentrations of AcTyr and AcTrp solution and found that the excited-state lifetime of Atto655/AcTyr complex is about 200 times longer than that of Atto655/AcTrp complex.