• Title/Summary/Keyword: fluorescence lifetime

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PROTEIN CONFORMATIONS OF OCTOPUS RHODOPSIN AND ITS DEPROTONATED PHOTOCYCLE INTERMEDIATE MONITORED BY ABSORPTION AND PROTEIN FLUORESCENCE

  • Jang, Du-Jeon;Lee, SunBae
    • Journal of Photoscience
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    • v.2 no.1
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    • pp.19-25
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    • 1995
  • Picosecond time-resolved and static protein fluorescence spectra and absorption spectra of octopus rhodopsin, a photorecepting protein, are measured and compared with those of bacteriorhodopsin, a photon-induced proton pumping protein, to understand the protein conformations and functions of octopus rhodopsin and its deprotonated photocycle intermediate. The bluer and weaker absorption of retinal indicates that octopus rhodopsin is better in thermal noise suppression but less efficient in light harvesting than bacteriorhodopsin. The protein fluorescence of octopus rhodopsin shows the characteristic of Trp only and the uantum efficiency and lifetime variations may result primarily from variations in the coupling strength with the retinal. The stronger intensity by four times and larger red shift by 12 nm of fluorescence suggest that octopus rhodopsin has more open and looser structure compared with bacteriorhodopsin. Fluorescence decay profiles reveal two decay components of 300 ps (60%) and 2 ns (40%). The deprotonation of protonated Schiff's base increases the shorter decay time to 500 ps and enhances the fluorescence intensity by 20%. The fluorescence and its decay time from Trp residues near retinal are influenced more by the deprotonation. The increase of fluorescence intimates that protein structure becomes loosened and relaxed further by the deprotonation of protonated Schiff's base. The driving force of sequential changes initiated by absorption of a photon is too exhausted after the deprotonation to return the intermediate to the ground state of the begun rhodopsin form.

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Photochemical synthesis of diphenylphenanthrenes, and the photophysical properties studied by emission and transient absorption measurements

  • Yamaji, Minoru;Hakoda, Yuuma;Horimoto, Ami;Okamoto, Hideki
    • Rapid Communication in Photoscience
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    • v.3 no.4
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    • pp.73-75
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    • 2014
  • Novel diphenylphenanthrenes (DPPs) were prepared by a photocyclization method, and the substituent effects of the phenyl groups on the photophysical properties of the phenanthrene chromophore were investigated based on measurements of fluorescence yields, lifetimes, and transient absorption. Fluorescence activities in DPPs are increased by introducing phenyl rings that can enhance the transition moment along the short axis of the phenanthrene skeleton. Intersystem crossing from the fluorescent states to the triplet manifolds is shown to be operative through the triplet-triplet absorption spectra obtained by laser photolysis techniques.

PHOTOPHYSICAL AND OPTICAL PROBE PROPERTIES OF 1-(p-N,N-DIMETHYLAMINOPHENYL)-4-PHENYL-2-METHYL-1E,3E-BUTADIENE

  • Singh, A.K.;Krishna, T.S.R.
    • Journal of Photoscience
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    • v.4 no.1
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    • pp.1-5
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    • 1997
  • A hitherto unknown diphenylbutadiene analog viz. 1-(p-N,N-dimethylaminophenyl)-4-phenyl-2-methyl-1E,3E-butadiene (10) has been prepared and its absorption, excitation, and fluorescent emission properties in different media including various organic solvents and aqueous bovine serum albumin (BSA) have been studied. For comparision, these properties have also been investigated for the parent diphenylbutadiene (2). Diene 10 exhibits solvent polarity/polarizability-sensitive fluorescence properties ($\lambda$$_{max}$, $\Phi$$_f$, $\tau$$_f$, K$_f$, f). It also binds to the hydrophobic domains of aqueous bovine serum albumin (BSA) with a binding constant of 3.89 x 10$^4$ M$^{-1}$. The relative fluorescence quantum yield of 10 increases, while, the fluorescence lifetime decreases with increasing concentration of-BSA. The results highlight the polar character of the singlet excited state of diphenylpolyenes and the utility of 10 as fluorescence probe for studying microenvironments of organized assemblies and biological supramolecular structures.

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Effect of Deuterated Solvents on the Excited State Photophysical Properties of Curcumin

  • Barik, A.;Goel, N.K.;Priyadarsini, K.I.;Mohan, Hari
    • Journal of Photoscience
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    • v.11 no.3
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    • pp.95-99
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    • 2004
  • Optical absorption and emission studies have been carried out to understand the effect of deuterium on the solvent dependent photophysical properties of curcumin in deuterated solvents such as $CDCl_3,\;(CD_3)_2SO,\;(CD_3)_2CO,\;CD_3OD\;and\;CD_3CN$. Optical absorption spectral studies showed that there is no significant shift in absorption maxima compared to the non-deuterated solvent. The fluorescence maxima shows significant shift with polarity of solvent but not much affected by the deuteration. The fluorescence quantum yield of curcumin increased marginally in almost all the deuterated solvents, indicating reduction in the non-radiative pathways. The fluorescence decay was biexponential in all the solvents and the average fluorescence lifetime was not much affected with deuteration, but showed decrease with increasing solvent polarity. Based on these studies, it is concluded that intermolecular hydrogen transfer is only partially responsible for the excited state deactivation of curcumin.

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Effects of Light Pulse Intensity and Quencher Concentration on the Time-Dependent Fluorescence Quenching Kinetics

  • Yang Mino;Lee Sangyoub;Shin, Kook Joe;Choo Kwang Yul;Lee Duckhwan
    • Bulletin of the Korean Chemical Society
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    • v.13 no.3
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    • pp.325-331
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    • 1992
  • By using the general theoretical framework proposed recently for treating the fluorescence quenching kinetics, we investigate the effect of light pulse intensity on the decay of fluorescence which follows excitation of fluorophors by the light pulse of very short but finite duration. It is seen that conventional theory breaks down when the exciting light pulse has a pulse width comparable to the fluorescent lifetime and its intensity is very high. We also find that even when the light intensity is not too high, conventional theory may fail in either of the following cases: (i) when the quencher concentration is high, (ii) when there is an attractive potential of mean force between the fluorophor and quencher, or (iii) when the energy transfer from the fluorophor to the quencher may also occur at a distance, e.g., via dipole-dipole interaction. The validity of the predictions of the present theory may thus be tested by fluorescence quenching experiments performed under such situations.

A FRET Assay for Celiac Disease

  • Lee, Sae A;Cho, Chul Min;Jang, Il Ho;Kang, Jung Sook
    • Biomedical Science Letters
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    • v.22 no.4
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    • pp.160-166
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    • 2016
  • To provide a basis for a homogeneous fluorescence resonance energy transfer (FRET) immunoassay for celiac disease, we carried out a FRET experiment using guinea pig tissue transglutaminase (tTG) and antibodies to tTG (anti-tTG) purified from rat serum. Fluorescein was utilized as the probe, and a nonfluorescent dye, QSY 7 served as the quencher. We labeled anti-tTG and tTG with fluorescein isothiocyanate and QSY 7 succinimidyl ester, respectively. Fluorescein-labeled anti-tTG was the donor, and QSY 7-labeled tTG was the acceptor of the FRET experiment. When we titrated fluorescein-labeled anti-tTG with QSY 7-labeled tTG, we observed a large decrease in the steady-state fluorescence intensity, which was due to strong FRET from fluorescein-labeled anti-tTG to QSY 7-labeled tTG. Using time-resolved fluorescence spectroscopy, we could also observe a decrease in the fluorescence lifetime, which confirms the steady-state data. We expect that these results might be useful in the development of a novel fluorescence immunoassay for an easy screening and follow-up of celiac patients.