title: [A method for determining the individual optical characteristics of posttranslational fluorescent forms of fluorescent proteins with the use of nonlinear laser fluorimetry].
abstract: A method for determining the individual optical characteristics (fluorescence quantum yield, the rate constant and quantum yield of singlet-triplet conversion, excitation of fluorescence cross-section, extinction coefficient) and concentration correlations between the fluorescent forms of fluorescent proteins arising in the reaction of posttranslational chromophore formation has been developed, which is based on combined application of absorption spectroscopy and classical and nonlinear laser fluorimretry. The method allows one to determine the share of fluorescent forms in the mixture of chromoproteins. The individual optical characteristics of the red form of the fluorescent protein mRFP1 has been determined: the fluorescence quantum yield eta = 0.24 +/- 0.03; the extinction coefficient in the maximum of absorbance band (584 nm) epsilon = 213 +/- 40 mM(-1) cm(-1) (the cross-section of absorbance sigma = (8.2 +/- 1.5).10(-16) cm2); the constant of singlet-triplet conversion rate K32 = (0 +/- 0.6)-10970 s(-1). The part of the red form in the mixture of chromoproteins is 26 +/- 6%.
authors.forename: A A
authors.initials: AA
authors.lastname: Banishev
authors.forename: E P
authors.initials: EP
authors.lastname: Vrzhets
authors.forename: D V
authors.initials: DV
authors.lastname: Dmitrienko
authors.forename: V L
authors.initials: VL
authors.lastname: Drutsa
authors.forename: D V
authors.initials: DV
authors.lastname: Maslov
authors.forename: V Z
authors.initials: VZ
authors.lastname: Pashchenko
authors.forename: E A
authors.initials: EA
authors.lastname: Shirshin
authors.forename: P V
authors.initials: PV
authors.lastname: Vrzhets
authors.forename: V V
authors.initials: VV
authors.lastname: Fadeev
mesh_terms: D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000737:chemistry*; D007834:Lasers; D014176:Protein Biosynthesis; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry*; D013050:Spectrometry, Fluorescence
chemical_list: D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins