scholarly journals Interaction of Zn2+ and Eu3+ with bovine liver glutamate dehydrogenase

1987 ◽  
Vol 246 (1) ◽  
pp. 199-203 ◽  
Author(s):  
E T Bell ◽  
A M Stilwell ◽  
J E Bell
Keyword(s):  
Binding Site ◽  
Glutamate Dehydrogenase ◽  
Quaternary Structure ◽  
Resonance Energy Transfer ◽  
Fluorescence Emission ◽  
Resonance Energy ◽  
Bovine Liver ◽  
Heat Stability ◽  
Protein Fluorescence ◽  
Excitation Peak

Bovine liver glutamate dehydrogenase is potently inhibited by Zn2+ ions. At pH 7.0 a kinetic dissociation constant for Zn2+ of 18 microM is obtained. The fluorescent lanthanide Eu3+ competes for the Zn2+-binding site and relieves the Zn2+-induced inhibition, but does not cause inhibition. Studies on the effects of Zn2+ or Eu3+ on the tertiary and quaternary structure of the enzyme by the use of protein fluorescence, heat-stability and re-activation after guanidinium chloride denaturation indicate that, whereas Zn2+ affects both tertiary and quaternary structure, Eu3+ does not affect either, consistent with its lack of effect on enzymic properties. Eu3+ fluorescence had a strong excitation peak at 395 nm with emission at 456 nm. In the presence of glutamate dehydrogenase the fluorescence emission is shifted to 501 nm. Eu3+, with high-affinity binding site and distinctive fluorescence properties after binding, would appear to be an ideal fluorophore for use in conformational studies or resonance-energy-transfer studies.

scholarly journals Preparation of CdTe/Alginate Textile Fibres with Controllable Fluorescence Emission through a Wet-Spinning Process and Application in the Trace Detection of Hg2+ Ions

Nanomaterials ◽  
2019 ◽  
Vol 9 (4) ◽  
pp. 570 ◽  
Author(s):  
Zhihui Zhao ◽  
Cunzhen Geng ◽  
Xihui Zhao ◽  
Zhixin Xue ◽  
Fengyu Quan ◽  
...  
Keyword(s):  
Mechanical Strength ◽  
Resonance Energy Transfer ◽  
Fluorescence Emission ◽  
Resonance Energy ◽  
Coagulation Bath ◽  
Wet Spinning ◽  
Trace Detection ◽  
Metallic Ions ◽  
Textile Fibres ◽  
Cdte Nanocrystals

Fluorescent textile fibres (FTFs) are widely used in many industrial fields. However, in addition to fibres with good fluorescence, fibres with excellent colour controllability, structural stability and appropriate mechanical strength still need to be developed. In this work, CdTe/alginate composite FTFs are prepared by taking advantage of the interactions between CdTe nanocrystals (NCs) and alginate macromolecules via a wet-spinning machine with a CaCl2 aqueous solution as the coagulation bath. CdTe NCs were chemically fixed in the fibre due to the interactions among surface ligands, macromolecules and coagulators (calcium ions), which ensured the excellent dispersity and good stability of the fibres. Förster resonance energy transfer (FRET) between NCs in the fibre was found to be restricted, which means that the emission colour of the fibres was totally controllable and could be predicted. Other properties of alginate fibres, such as flame retardance and mechanical strength, were also well preserved in the fluorescent fibres. Finally, FTFs showed good selectivity toward trace Hg2+ ions over other metallic ions, and the detection could be identified by the naked eye.

scholarly journals A quantitative protocol for dynamic measurements of protein interactions by Förster resonance energy transfer-sensitized fluorescence emission

2008 ◽  
Vol 6 (suppl_1) ◽  
Author(s):  
A.D Elder ◽  
A Domin ◽  
G.S Kaminski Schierle ◽  
C Lindon ◽  
J Pines ◽  
...  
Keyword(s):  
Energy Transfer ◽  
Protein Interactions ◽  
Imaging Techniques ◽  
Resonance Energy Transfer ◽  
Fluorescence Emission ◽  
Resonance Energy ◽  
Forster Resonance Energy Transfer ◽  
Confocal Imaging ◽  
Förster Resonance Energy Transfer ◽  
Förster Resonance

Fluorescence detection of acceptor molecules sensitized by Förster resonance energy transfer (FRET) is a powerful method to study protein interactions in living cells. The method requires correction for donor spectral bleed-through and acceptor cross-excitation as well as the correct normalization of signals to account for varying fluorophore concentrations and imaging parameters. In this paper, we review different methods for FRET signal normalization and then present a rigorous model for sensitized emission measurements, which is both intuitive to understand and practical to apply. The method is validated by comparison with the acceptor photobleaching and donor lifetime-imaging techniques in live cell samples containing EYFP and ECFP tandem constructs exhibiting known amounts of FRET. By varying the stoichiometry of interaction in a controlled fashion, we show that information on the fractions of interacting donors and acceptors can be recovered. Furthermore, the method is tested by performing measurements on different microscopy platforms in both widefield and confocal imaging modes to show that signals recovered under different imaging conditions are in quantitative agreement. Finally, the method is applied in the study of dynamic interactions in the cyclin–cdk family of proteins in live cells. By normalizing the obtained signals for both acceptor and donor concentrations and using a FRET exhibiting control construct for calibration, stoichiometric changes in these interactions could be visualized in real time. The paper is written to be of practical use to researchers interested in performing sensitized emission measurements. The correct interpretation of the retrieved signals in a biological context is emphasized, and guidelines are given for the practical application of the developed algorithms.

scholarly journals Bioluminescence-Based Energy Transfer Using Semiconductor Quantum Dots as Acceptors

Sensors ◽  
2020 ◽  
Vol 20 (10) ◽  
pp. 2909 ◽  
Author(s):  
Anirban Samanta ◽  
Igor L. Medintz
Keyword(s):  
Quantum Dots ◽  
Energy Transfer ◽  
Organic Dyes ◽  
Stokes Shift ◽  
Resonance Energy Transfer ◽  
Fluorescence Emission ◽  
Resonance Energy ◽  
Semiconductor Quantum Dots ◽  
Renilla Luciferase ◽  
Bioluminescent Protein

Bioluminescence resonance energy transfer (BRET) is the non-radiative transfer of energy from a bioluminescent protein donor to a fluorophore acceptor. It shares all the formalism of Förster resonance energy transfer (FRET) but differs in one key aspect: that the excited donor here is produced by biochemical means and not by an external illumination. Often the choice of BRET source is the bioluminescent protein Renilla luciferase, which catalyzes the oxidation of a substrate, typically coelenterazine, producing an oxidized product in its electronic excited state that, in turn, couples with a proximal fluorophore resulting in a fluorescence emission from the acceptor. The acceptors pertinent to this discussion are semiconductor quantum dots (QDs), which offer some unrivalled photophysical properties. Amongst other advantages, the QD’s large Stokes shift is particularly advantageous as it allows easy and accurate deconstruction of acceptor signal, which is difficult to attain using organic dyes or fluorescent proteins. QD-BRET systems are gaining popularity in non-invasive bioimaging and as probes for biosensing as they don’t require external optical illumination, which dramatically improves the signal-to-noise ratio by avoiding background auto-fluorescence. Despite the additional advantages such systems offer, there are challenges lying ahead that need to be addressed before they are utilized for translational types of research.

Location of the stilbenedisulfonate binding site of the human erythrocyte anion-exchange system by resonance energy transfer

Biochemistry ◽  
1979 ◽  
Vol 18 (21) ◽  
pp. 4505-4516 ◽  
Author(s):  
Anjana Rao ◽  
Paul Martin ◽  
Reinhart A. F. Reithmeier ◽  
Lewis C. Cantley
Keyword(s):  
Energy Transfer ◽  
Binding Site ◽  
Anion Exchange ◽  
Human Erythrocyte ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Exchange System

scholarly journals Quaternary Structure Assessment of ICln by Fluorescence Resonance Energy Transfer (FRET) in vivo

2009 ◽  
Vol 23 (4-6) ◽  
pp. 397-406 ◽  
Author(s):  
Sabine Schmidt ◽  
Martin Jakab ◽  
Ivano Costa ◽  
Johannes Fürst ◽  
Andrea Ravasio ◽  
...  
Keyword(s):  
Energy Transfer ◽  
Fluorescence Resonance Energy Transfer ◽  
Quaternary Structure ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Structure Assessment ◽  
Fluorescence Resonance

Understanding of Förster Resonance Energy Transfer (FRET) in Ionic Materials

2021 ◽  
Vol 2 (4) ◽  
pp. 564-575
Author(s):  
Amanda Jalihal ◽  
Thuy Le ◽  
Samantha Macchi ◽  
Hannah Krehbiel ◽  
Mujeebat Bashiru ◽  
...  
Keyword(s):  
Energy Transfer ◽  
Nile Blue ◽  
Photophysical Properties ◽  
Resonance Energy Transfer ◽  
Fluorescence Emission ◽  
Resonance Energy ◽  
Forster Resonance Energy Transfer ◽  
Förster Resonance Energy Transfer ◽  
Fluorescence Emission Spectrum ◽  
Förster Resonance

Herein, an ionic material (IM) with Förster Resonance Energy Transfer (FRET) characteristics is reported for the first time. The IM is designed by pairing a Nile Blue A cation (NBA+) with an anionic near-infrared (NIR) dye, IR820−, using a facile ion exchange reaction. These two dyes absorb at different wavelength regions. In addition, NBA+ fluorescence emission spectrum overlaps with IR820− absorption spectrum, which is one requirement for the occurrence of the FRET phenomenon. Therefore, the photophysical properties of the IM were studied in detail to investigate the FRET mechanism in IM for potential dye sensitized solar cell (DSSCs) application. Detailed examination of photophysical properties of parent compounds, a mixture of the parent compounds, and the IM revealed that the IM exhibits FRET characteristics, but not the mixture of two dyes. The presence of spectator counterion in the mixture hindered the FRET mechanism while in the IM, both dyes are in close proximity as an ion pair, thus exhibiting FRET. All FRET parameters such as spectral overlap integral, Förster distance, and FRET energy confirm the FRET characteristics of the IM. This article presents a simple synthesis of a compound with FRET properties which can be further used for a variety of applications.

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