Intrinsic fluorescence as a probe of structure-function relationships in Ca2+-transport ATPases

1996 ◽  
Vol 16 (2) ◽  
pp. 87-106 ◽  
Author(s):  
Sérgio T. Ferreira ◽  
Tatiana Coelho-Sampaio
Keyword(s):  
Structure Function ◽  
Conformational Changes ◽  
Fluorescence Emission ◽  
Intrinsic Fluorescence ◽  
Future Prospects ◽  
Time Resolved Fluorescence ◽  
Time Resolved ◽  
Fluorescence Measurements ◽  
Transport Atpases ◽  
Functional States

Applications of intrinsic fluorescence measurements in the study of Ca2+-transport ATPases are reviewed. Since the initial reports showing that the fluorescence emission was sensitive to Ca2+ binding, a substantial amount of work has focused on the use of both steady-state and time-resolved fluorescence spectroscopy to investigate structure-function relationships in sarcoplasmic reticulum and plasma membrane Ca2+-ATPases. These studies have revealed ligand-induced conformational changes, as well as provided information on protein-protein, protein-solvent and/or protein-lipid interactions in different functional states of these proteins. The main results of these studies, as well as possible future prospects are discussed.

Fluorescence study on transmembrane Ca2+ gradient-mediated conformation changes of sarcoplasmic reticulum Ca2+-ATPase

1994 ◽  
Vol 14 (6) ◽  
pp. 309-317 ◽  
Author(s):  
Y. P. Tu ◽  
F. Y. Yang
Keyword(s):  
Sarcoplasmic Reticulum ◽  
Fluorescence Quenching ◽  
Conformational Changes ◽  
Dynamic Change ◽  
Parasitic Fungus ◽  
Time Resolved Fluorescence ◽  
Fluorescence Study ◽  
Time Resolved ◽  
Fluorescence Measurements ◽  
Tryptophan Residues

The conformational states of Ca2+-ATPase in sarcoplasmic reticulum (SR) vesicles with or without a thousand-fold transmembrane Ca2+ gradient have been studied by fluorescence spectroscopy and fluorescence quenching. In consequence of the establishment of the transmembrane Ca2+ gradient, the steady-state fluorescence results revealed a reproducible 8% decrease in the intrinsic fluorescence while time-resolved fluorescence measurements showed that 13 tryptophan residues in SR · Ca2+-ATPase could be divided into three groups. The fluorescence lifetime of one of these groups increased from 5.5 ns to 5.95 ns in the presence of a Ca2+ gradient. Using KI and hypocrellin B (a photosensitive pigment obtained from a parasitic fungus, growing in Yunnan, China), the fluorescence quenching further indicated that the dynamic change of this tryptophan group, located at the protein-lipid interface, is a characteristic of transmembrane Ca2+ gradient-mediated conformational changes in SR · Ca2+-ATPase.

Sensitive, rapid procedure for time-resolved immunofluorometry of lutropin.

1988 ◽  
Vol 34 (8) ◽  
pp. 1640-1644 ◽  
Author(s):  
M J Khosravi ◽  
R C Morton ◽  
E P Diamandis
Keyword(s):  
Monoclonal Antibody ◽  
Solid Phase ◽  
Detection System ◽  
Time Resolved Fluorescence ◽  
Time Resolved ◽  
Practical Procedure ◽  
Daily Monitoring ◽  
Fluorescence Measurements ◽  
Capture Antibody ◽  
Europium Chelate

Abstract In this new immunofluorometric method for quantification of lutropin in serum, the "sandwich" principle is combined with time-resolved fluorescence measurements, with the europium chelate 4,7-bis(chlorosulfophenyl)-1,10-phenanthroline-2,9-dicarboxylic acid (BCPDA) used as label. A monoclonal antibody to the alpha-subunit of lutropin is adsorbed onto the walls of white-opaque microtiter wells to form the solid-phase capture antibody, and a biotin-labeled soluble monoclonal antibody is used for antigen quantification. The detection system is completed with streptavidin, which has been linked to a protein bulking agent labeled with multiple BCPDA residues. In the presence of excess europium, the fluorescence of the final complex attached to captured lutropin molecules is measured on the dried solid phasse with an automated time-resolved fluorometer. The assay can be performed as a rapid (less than 60 min incubation) or regular (150 min incubation) procedure. The rapid assay is well-suited for routine daily monitoring of increasing or ovulatory lutropin concentrations; the regular assay, with its greater sensitivity (0.5 int. unit/L), is a practical procedure for lutropin measurements in hyposecretory states. The assay measures up to 240 int. units/L, and results compare well with those by a commercially available radioimmunoassay, an immunoradiometric assay, and another time-resolved immunofluorometric procedure.

FRET Study Between Carbon Quantum Dots and Malachite Green by Steady-State and Time-Resolved Fluorescence Spectroscopy

2020 ◽  
Vol 10 (3) ◽  
pp. 178-188
Author(s):  
Bipin Rooj ◽  
Ankita Dutta ◽  
Debojyoti Mukherjee ◽  
Sahidul Islam ◽  
Ujjwal Mandal
Keyword(s):  
Quantum Dots ◽  
Energy Transfer ◽  
Fluorescence Spectroscopy ◽  
Malachite Green ◽  
Dye Degradation ◽  
Fluorescence Emission ◽  
Carbon Quantum Dots ◽  
Time Resolved Fluorescence ◽  
Physical Processes ◽  
Time Resolved

Background: Understanding the interaction between different organic dyes and carbon quantum dots helps us to understand several photo physical processes like electron transfer, energy transfer, molecular sensing, drug delivery and dye degradation processes etc. Objective: The primary objective of this study is to whether the carbon quantum dots can act as an electron donor and can participate in the different photo physical processes. Methods: In this work, Carbon Quantum Dots (CQDLs) are synthesized in most economical and simple carbonization method where petals of Nelumbo nucifera L. are used as a carbon precursor. The synthesized CQDLs were characterized by using experimental techniques like UV−Vis absorption, FT-IR, Transmission Electron Microscopy (TEM), steadystate and time-resolved fluorescence spectroscopy. Results: The spectral analysis shows that the so synthesized CQDLs are spherical in shape and its diameter is around 4.2 nm. It shows the fluorescence emission maximum at 495 nm with a quantum yield of 4%. In this work the interaction between Carbon Quantum Dots (CQDLs) and an organic dye Malachite Green (MG) is studied using fluorescence spectroscopic technique under ambient pH condition (At pH 7). The quenching mechanism of CQDLs with MG was investigated using Stern-Volmer equation and time-resolved fluorescence lifetime studies. The results show that the dominant process of fluorescence quenching is attributed to Forster Resonance Energy Transfer (FRET) having a donor acceptor distance of 53 Å where CQDLs act as a donor and MG acts as an acceptor. Conclusion: This work has a consequence that CQDLs can be used as a donor species for different photo physical processes such as photovoltaic cell, dye sensitized solar cell, and also for antioxidant activity study.

scholarly journals Development, Characterization, and Application of a Versatile Single Particle Detection Apparatus for Time-Integrated and Time-Resolved Fluorescence Measurements—Part II: Experimental Evaluation

2009 ◽  
Vol 2009 ◽  
pp. 1-14
Author(s):  
Xihong Wu ◽  
J. A. Merten ◽  
N. Omenetto ◽  
B. W. Smith ◽  
J. D. Winefordner
Keyword(s):  
Single Particle ◽  
Narrow Beam ◽  
Time Resolved Fluorescence ◽  
Particle Detection ◽  
Time Resolved ◽  
Endogenous Fluorophores ◽  
Fluorescence Measurements ◽  
On Line ◽  
Single Particle Detection ◽  
Speed Mode

This paper describes the experimental realization and characterization of a versatile single particle detection apparatus. The system utilizes a novel particle beam inlet that can serve as either an on-line particle concentrator (i.e., all diameters confined in a narrow beam) or as a segregator (i.e., selected diameters confined in a narrow beam) and can be operated in a high-speed mode as well as in a low-speed mode, thus allowing different interaction times between the particles and the laser beam. An aerodynamic sizing technique has been incorporated into the system to provide rapid, real-time, and high-resolution sizing. Parameters such as transmission efficiency and size-segregation efficiency have been measured. The performance of the instrument has been demonstrated by on-line detection of spectrally resolved and time resolved fluorescence detection from airborne dye-doped particles and aerosolized endogenous fluorophores found in biological agents.

Fluorescence property of photosystem II protein complexes bound to a gold nanoparticle

2017 ◽  
Vol 198 ◽  
pp. 121-134 ◽  
Author(s):  
Kazuki Tahara ◽  
Ahmed Mohamed ◽  
Kousuke Kawahara ◽  
Ryo Nagao ◽  
Yuki Kato ◽  
...  
Keyword(s):  
Photosystem Ii ◽  
Gold Nanoparticle ◽  
Water Oxidation ◽  
Artificial Photosynthesis ◽  
Fluorescence Property ◽  
Light Illumination ◽  
Time Resolved Fluorescence ◽  
Strong Light ◽  
Time Resolved ◽  
Fluorescence Measurements

Development of an efficient photo-anode system for water oxidation is key to the success of artificial photosynthesis. We previously assembled photosystem II (PSII) proteins, which are an efficient natural photocatalyst for water oxidation, on a gold nanoparticle (GNP) to prepare a PSII–GNP conjugate as an anode system in a light-driven water-splitting nano-device (Noji et al., J. Phys. Chem. Lett., 2011, 2, 2448–2452). In the current study, we characterized the fluorescence property of the PSII–GNP conjugate by static and time-resolved fluorescence measurements, and compared with that of free PSII proteins. It was shown that in a static fluorescence spectrum measured at 77 K, the amplitude of a major peak at 683 nm was significantly reduced and a red shoulder at 693 nm disappeared in PSII–GNP. Time-resolved fluorescence measurements showed that picosecond components at 683 nm decayed faster by factors of 1.4–2.1 in PSII–GNP than in free PSII, explaining the observed quenching of the major fluorescence peak. In addition, a nanosecond-decay component arising from a ‘red chlorophyll’ at 693 nm was lost in time-resolved fluorescence of PSII–GNP, probably due to a structural perturbation of this chlorophyll by interaction with GNP. Consistently with these fluorescence properties, degradation of PSII during strong-light illumination was two times slower in PSII–GNP than in free PSII. The enhanced durability of PSII is an advantageous property of the PSII–GNP conjugate in the development of an artificial photosynthesis device.

scholarly journals Effect of the Substitution Position on the Electronic and Solvatochromic Properties of Isocyanoaminonaphthalene (ICAN) Fluorophores

Molecules ◽  
2019 ◽  
Vol 24 (13) ◽  
pp. 2434 ◽  
Author(s):  
Sándor Lajos Kovács ◽  
Miklós Nagy ◽  
Péter Pál Fehér ◽  
Miklós Zsuga ◽  
Sándor Kéki
Keyword(s):  
Radiative Decay ◽  
Fluorescence Emission ◽  
Solvent Polarity ◽  
Decay Rates ◽  
Quantum Yields ◽  
Time Resolved Fluorescence ◽  
Solvatochromic Shift ◽  
Fluorescence Emission Spectra ◽  
Time Resolved ◽  
Donor And Acceptor

The properties of 1,4-isocyanoaminonaphthalene (1,4-ICAN) and 2,6-isocyanoaminonaphthalene (2,6-ICAN) isomers are discussed in comparison with those of 1,5-isocyanoaminonaphthalene (1,5-ICAN), which exhibits a large positive solvatochromic shift similar to that of Prodan. In these isocyanoaminonaphthalene derivatives, the isocyano and the amine group serve as the donor and acceptor moieties, respectively. It was found that the positions of the donor and the acceptor groups in these naphthalene derivatives greatly influence the Stokes and solvatochromic shifts, which decrease in the following order: 1,5-ICAN > 2,6-ICAN > 1,4-ICAN. According to high-level quantum chemical calculations, this order is well correlated with the charge transfer character of these compounds upon excitation. Furthermore, unlike 1,5-ICAN, the 1,4-ICAN and 2,6-ICAN isomers showed relatively high quantum yields in water, that were determined to be 0.62 and 0.21, respectively. In addition, time-resolved fluorescence experiments revealed that both the radiative and non-radiative decay rates for these three ICAN isomers varied unusually with the solvent polarity parameter ET(30). The explanations of the influence of the solvent polarity on the resulting steady-state and time-resolved fluorescence emission spectra are also discussed.

An integrated instrumentation for light‐scattering and time‐resolved fluorescence measurements

1995 ◽  
Vol 66 (3) ◽  
pp. 2405-2410 ◽  
Author(s):  
M. Musolino ◽  
R. Cubeddu ◽  
A. Pifferi ◽  
P. Taroni ◽  
P. Lago ◽  
...  
Keyword(s):  
Light Scattering ◽  
Time Resolved Fluorescence ◽  
Time Resolved ◽  
Fluorescence Measurements
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