The fluorescence decay of human serum albumin and its subfractions

Hazan, Gershon; Haas, Elisha; Steinberg, Izchak Z.

doi:10.1016/0005-2795(76)90044-1

Cited by 31 publications

(16 citation statements)

References 12 publications

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“…DeLauder and Wahl [5], for example, found two components (in charcoal treated HSA at pH 5.5) of 3.3 ns (66%) and 7.8 ns (34%) and Wahl and Auchet [6]later analyzed the decay as a function of the emission wavelength and, upon 295 nm excitation, found three components, namely, 1.5 ns, 6.17 ns and 12.08 ns, with fractional contributions dependent upon the emission wavelength but with 6.17 ns being by far the major component. Hazan et al [7]resolved the decay of HSA, upon 296 nm excitation and at pH 7.4, into two exponentials having lifetimes of 6.1 ns (45%) and 1.5 ns (55%) while Kasai et al [8]found two components of 6.89 ns (46.5%) and 3.22 ns (53.5%) using 295 nm excitation at pH 7.4. Using multifrequency phase and modulation fluorometry, Lakowicz and Gryczynski [13]found for HSA, upon excitation at 298 nm at 20°C and pH 8, two decay components of 6.06 ns (95.7%) and 1.42 ns (4.3%).…”

Section: Discussionmentioning

confidence: 99%

“…The excited state properties of Trp‐214, including both lifetime and rotational properties, have been studied for normal HSA [4–14]. The average lifetime of Trp‐214 in FDH‐HSA was reported to be less than in normal HSA [15], but was difficult to quantify since the data were obtained on proteins isolated from persons heterozygous for FDH‐HSA, i.e.…”

Section: Introductionmentioning

confidence: 99%

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Time‐resolved fluorescence studies on site‐directed mutants of human serum albumin

et al. 1997

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Time‐resolved fluorescence studies on site‐directed mutants of human serum albumin

et al. 1997

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“…The fraction (f) of Trp residues that emits sufficiently to be recorded (Ricci 1970;Hazen et al 1976) is given by,…”

Section: The Stern-volmer Data For the Cs § And T1 § Quenching Of Thementioning

confidence: 99%

A differential molecualr topography of the Pr and Pfr forms of native oat phytochrome as probed by fluoresence quenching

Singh

Song

1990

Planta

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Tryptophan (Trp) surface topography of the red- and far-red-absorbing forms of phytochrome (Pr, Pfr) ofAvena sativa L. has been investigated by analyzing quenching of the two components of Trp fluorescence decay, in order to understand the differences in the two forms at the molecular level. Stern-Volmer kinetic analysis of the quenching data for two cationic surface quenchers, Cs(+) and Tl(+), showed strong quenching of the short component of the Pr fluorescence (Stern-Volmer constants,K sv , 27.2 and 21.4 M(-1), respectively) relative to that of Pfr fluorescenceK sv , 10.4 and 12.3 M(-1), respectively). The long component of the Trp fluorescence was quenched differentially by Cs(+) and Tl(+), withK sv of 9.0 and 19.8 M(-1), respectively, for the Pr fluorescence andK sv of 13.7 and 8.7 M(-1), respectively, for the Pfr fluorescence. The results indicate that the phytochrome Trp residues with short fluorescence lifetime are more accessible to the cationic surface quenchers than those with long fluorescence lifetime. The data, taken together with our earlier study (Singh et al. 1988, Biochim, Biophys. Acta936, 395-405), indicate that most, if not all the ten Trp residues of phytochrome, are fluorescent and exist in distinct groups differing in their topography and microenvironment, and the peptide segment containing Trp-774 and Trp-778 within the 55-kilodalton C-terminal domain of phytochrome also undergoes a subtle alteration in its surface topography during Pr→Pfr phototransformation.

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“…4B). Hazan et al [47] also showed that monomeric and dimeric HSA have different excited state lifetime properties. This difference underscores the necessity of isolating monomeric HSA from the aggregates usually found in commercial lyophilized preparations before undertaking a study of its intrinsic tryptophan fluorescence, an obvious step that, surprisingly, is sometimes overlooked.…”

Section: Resultsmentioning

confidence: 99%

Applications of phasor plots to in vitro protein studies

James

Ross

Štefl

et al. 2011

Analytical Biochemistry

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The previous paper describes the application of the phasor analysis to fluorescence intensity decay data on in vitro samples. As detailed in that paper, this method provides researchers with a simple graphical method for viewing lifetime data that can be used to quantify individual components of a mixture as well as to identify excited state reactions. In this report, we extend the use of in vitro phasor analysis to intrinsic protein fluorescence. We show how alterations in the excited state properties of tryptophan residues are easily visualized using the phasor method. Specifically, we demonstrate that protein-ligand and protein-protein interactions can result in unique shifts in the location of phasor points, indicative of protein conformational changes. Application of the method to a rapid kinetics experiment is also shown. Finally, we show that the unfolding of lysozyme with either urea or guanidine hydrochloride results in different phasor trajectories, indicative of unique denaturation pathways.

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The fluorescence decay of human serum albumin and its subfractions

Cited by 31 publications

References 12 publications

Time‐resolved fluorescence studies on site‐directed mutants of human serum albumin

Time‐resolved fluorescence studies on site‐directed mutants of human serum albumin

A differential molecualr topography of the Pr and Pfr forms of native oat phytochrome as probed by fluoresence quenching

Applications of phasor plots to in vitro protein studies

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