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Forster resonance energy transfer

Jürgen M Plitzko, Benjamin Schuler, Philipp Selenko
Recent developments in cellular cryo-electron tomography, in-cell single-molecule Förster resonance energy transfer-spectroscopy, nuclear magnetic resonance-spectroscopy and electron paramagnetic resonance-spectroscopy delivered unprecedented insights into the inner workings of cells. Here, we review complementary aspects of these methods and provide an outlook toward joint applications in the future.
July 20, 2017: Current Opinion in Structural Biology
Nuala Del Piccolo, Kalina Hristova
Adaptor proteins are a class of cytoplasmic proteins that bind to phosphorylated residues in receptor tyrosine kinases and trigger signaling cascades that control critically important cellular processes, such as cell survival, growth, differentiation, and motility. Here, we seek to characterize the interaction between epidermal growth factor receptor (EGFR) and the cytoplasmic adaptor protein growth factor receptor-bound protein 2 (Grb2) in a cellular context. To do so, we explore the utility of a highly biologically relevant model system, mammalian cells under reversible osmotic stress, and a recently introduced Förster resonance energy transfer microscopy method, fully quantified spectral imaging...
July 19, 2017: Biophysical Journal
Weiqiang Yang, Jiancong Ni, Fang Luo, Wen Weng, Qiao-Hua Wei, Zhenyu Lin, Guonan Chen
Carbon dots (CDs) emerge as excellent fluorescent nanomaterials, but the full exploitation and application of their exceptional properties in the development of fluorescence assay are still rare. In this work, cationic carbon dots (C-CDs) covered with a plenty of positive charges on the surface were synthesized through a facile ultrasonic method. Negatively charged hyaluronic acid (HA) caused the aggregation of positively charged C-CDs and neutral red (NR) along its linear chain via electrostatic adsorption, leading to a remarkable förster resonance energy transfer (FRET) from C-CDs to NR...
July 21, 2017: Analytical Chemistry
Xiaoyu Huang, Heng Zhang, Dingxin Xu, Feng Wen, Shuming Chen
Exciton recombination zone, where the photons are generated, can greatly affect the performance such as the efficiency, the color purity of the quantum dot (QD) light-emitting diodes (QLEDs). To probe the exciton recombination zone, 4-(dicyanomethylene)-2-t-butyl-6(1,1,7,7 -tetramethyljulolidyl-9-enyl)-4H-pyran (DCJTB) is doped into the charge transport layer as a fluorescent sensor; by monitoring the Förster resonant energy transfer (FRET) between QD and DCJTB, the location of the recombination zone can be determined...
July 21, 2017: ACS Applied Materials & Interfaces
Moirangthem Kiran Singh, Mohammad Firoz Khan, Him Shweta, Sobhan Sen
Despite significant interest in understanding the role of the local dielectric environment and lipid-bilayer fluidity/rigidity in resonance energy transfer between chromophores at lipid/water interfaces, a comprehensive approach to quantify such environmental dependence on energy transfer is missing - primarily because of the scarcity of suitable probes. Here we present the results on multi-chromophoric Förster resonance energy transfer (FRET) from a series of 4-aminophthalimide-based molecules (4AP-Cn; n = 2-10, 12) of different lipophilicity (donors), which reside at different depths across the lipid/water interfaces, to rhodamine-6G (Rh6G; acceptor) molecules that stay in a water-rich region near the lipid headgroups...
July 20, 2017: Physical Chemistry Chemical Physics: PCCP
Yujin Zhang, Jiancai Leng
The development of fluorescent sensors for Hg(2+) has attracted much attention due to the well-known adverse effects of mercury on biological health. In the present work, the optical properties of two newly-synthesized Hg(2+) chemosensors based on the coumarin-rhodamine system (named Pro1 and Pro2) were systematically investigated using time-dependent density functional theory. It is shown that Pro1 and Pro2 are effective ratiometric fluorescent Hg(2+) probes, which recognize Hg(2+) by Förster resonance energy transfer and through bond energy transfer mechanisms, respectively...
July 20, 2017: Sensors
Gustavo Fuertes, Niccolò Banterle, Kiersten M Ruff, Aritra Chowdhury, Davide Mercadante, Christine Koehler, Michael Kachala, Gemma Estrada Girona, Sigrid Milles, Ankur Mishra, Patrick R Onck, Frauke Gräter, Santiago Esteban-Martín, Rohit V Pappu, Dmitri I Svergun, Edward A Lemke
Unfolded states of proteins and native states of intrinsically disordered proteins (IDPs) populate heterogeneous conformational ensembles in solution. The average sizes of these heterogeneous systems, quantified by the radius of gyration (RG ), can be measured by small-angle X-ray scattering (SAXS). Another parameter, the mean dye-to-dye distance (RE ) for proteins with fluorescently labeled termini, can be estimated using single-molecule Förster resonance energy transfer (smFRET). A number of studies have reported inconsistencies in inferences drawn from the two sets of measurements for the dimensions of unfolded proteins and IDPs in the absence of chemical denaturants...
July 17, 2017: Proceedings of the National Academy of Sciences of the United States of America
Alena Koukalová, Mariana Amaro, Gokcan Aydogan, Gerhard Gröbner, Philip T F Williamson, Ilya Mikhalyov, Martin Hof, Radek Šachl
It is a fundamental question in cell biology and biophysics whether sphingomyelin (SM)- and cholesterol (Chol)- driven nanodomains exist in living cells and in model membranes. Biophysical studies on model membranes revealed SM and Chol driven micrometer-sized liquid-ordered domains. Although the existence of such microdomains has not been proven for the plasma membrane, such lipid mixtures have been often used as a model system for 'rafts'. On the other hand, recent super resolution and single molecule results indicate that the plasma membrane might organize into nanocompartments...
July 14, 2017: Scientific Reports
Thomas-Otavio Peulen, Oleg Opanasyuk, Claus A M Seidel
Förster resonance energy transfer (FRET) measurements from a donor, D, to an acceptor, A, fluorophore are frequently used in vitro and in live cells to reveal information on the structure and dynamics of DA-labeled macromolecules. Accurate descriptions of FRET-measurements by molecular models are complicated because the fluorophores are usually coupled to the macromolecule via flexible long linkers allowing for diffusional exchange between multiple states with different fluorescence properties caused by distinct environmental quenching, dye mobilities, and variable DA-distances...
July 14, 2017: Journal of Physical Chemistry. B
Inka Negwer, Markus Hirsch, Stefka Kaloyanova, Tom Brown, Kalina Peneva, Hans-Jürgen Butt, Kaloian Koynov, Mark Helm
Cyanine dyes show a general propensity to localize to polarized mitochondria. This mitochondriotropism was used to perform a copper-free click reaction in the mitochondria of living cells. The in organello reaction of a Cy3 and Cy5 dye led to a product that was easily traceable by Förster resonance energy transfer (FRET). As determined by confocal laser scanning microscopy, the Cy3-Cy5 conjugate showed enhanced retention in mitochondria as compared to the starting compounds. This enhancement of a favorable property can be achieved by synthesis in organello but not outside mitochondria...
July 13, 2017: Chembiochem: a European Journal of Chemical Biology
Chang-Zheng Wang, Hai-Hao Han, Xin-Ying Tang, Dong-Ming Zhou, Changfeng Wu, Guo-Rong Chen, Xiao-Peng He, He Tian
The seasonal outbreak of influenza causes significant morbidity and mortality worldwide because a number of influenza virus (IV) strains have been shown to infect and circulate in humans. Development of effective means to timely monitor as well as block IVs is still a challenging task. Whereas conventional fluorescence probes rely on a fluorimetric change upon recognizing IVs, here we developed simple "Supra-dots" that are formed through the aqueous supramolecular assembly between a blue-emitting polymer dot and red-emitting sialylglycan probes for the ratiometric detection of IVs...
July 20, 2017: ACS Applied Materials & Interfaces
Sonashree Saxena, Manickam Jayakannan
The present investigation accounts one of the first example of enzyme-responsive and π-conjugate-tagged L-amino acid amphiphilic polymer and their fluorescence resonance energy transfer (FRET) probes for colour-tunable intracellular bio-imaging in cancer cells. Melt polymerizable oligo-phenylenevinylene (OPV) -conjugated diol was tailor-made and subjected to thermo-selective melt transesterification reaction with multi-functional L-aspartic acid monomer to yield OPV-tagged amphiphilic luminescent polyesters...
July 12, 2017: Biomacromolecules
Hans H Gorris, Ute Resch-Genger
In Part II of this review series on lanthanide-doped photon-upconversion nanoparticles (UCNPs), we present and critically discuss the performance and suitability of UCNPs as background-free luminescent reporters in bioimaging and bioanalytical applications. The preparation of a biocompatible nanoparticle surface is an integral step for all life - science-related applications. UCNPs have found their way into a large number of diagnostic platforms, homogeneous and heterogeneous assay formats, and sensor applications...
July 7, 2017: Analytical and Bioanalytical Chemistry
Muhammad Naufal Lintangpradipto, Nikolai Tsevtkov, Byeong Cheul Moon, Jeung Ku Kang
Here, we report that incorporation of size-controlled CdSe quantum dots (QDs) into perovskite photovoltaic cells (PSCs) boosts their light harvesting capability. X-ray photoemission and optical absorption spectroscopy analyses also show that the electronic structure of CdSe QDs makes them efficient charge transfer mediators between perovskite and Spiro-MeOTAD layers. In addition, electrochemical impedance spectroscopy experiments demonstrate that QDs help to decrease charge transfer resistance at the interfaces...
July 20, 2017: Nanoscale
Eleanna Kara, Jordan D Marks, Zhanyun Fan, Jacob A Klickstein, Allyson D Roe, Kelly A Krogh, Susanne Wegmann, Masato Maesako, Christina C Luo, Ravi Mylvaganam, Oksana Berezovska, Eloise Hudry, Bradley T Hyman
Apolipoprotein E (ApoE) has an important role in the pathogenesis of Alzheimer's disease with its three isoforms having distinct effects on disease risk. Here, we assessed the conformational differences between those isoforms using a novel flow cytometry-Forster Resonance Energy Transfer (FRET) assay. We showed that the conformation of intracellular ApoE within HEK cells and astrocytes adopts a directional pattern; in other words, E4 adopts the most closed conformation, E2 the most open conformation, and E3 an intermediate conformation...
July 6, 2017: Journal of Biological Chemistry
Jianhao Wang, Jie Fan, Li Liu, Shumin Ding, Xiaoqian Liu, Jianpeng Wang, Liqian Gao, Souvik Chattopadhaya, Peng Miao, Jiang Xia, Lin Qiu, Pengju Jiang
Herein, a novel assay has been developed for monitoring PreScission protease (His-PSP) mediated enzyme cleavage of ATTO 590 labeled peptide substrate (ATTO-LEV). This novel method is based on combining the use of capillary electrophoresis and fluorescence detection (CE-FL) to dynamically monitor the enzyme cleavage activity. A multivalent peptide substrate was first constructed by immobilizing His-tagged ATTO 590 labeled peptide substrate (ATTO-LEVH6) onto the surface of CdSe/ZnS quantum dots (QDs). Once successfully immobilized, the novel multivalent peptide substrate resulted in the Förster resonance energy transfer (FRET) from QDs to ATTO 590...
July 6, 2017: Electrophoresis
Elena Bollati, Daniel Plimmer, Cecilia D'Angelo, Jörg Wiedenmann
Photoconvertible fluorescent proteins (pcRFPs) are a group of fluorophores that undergo an irreversible green-to-red shift in emission colour upon irradiation with near-ultraviolet (near-UV) light. Despite their wide application in biotechnology, the high-level expression of pcRFPs in mesophotic and depth-generalist coral species currently lacks a biological explanation. Additionally, reduced penetration of near-UV wavelengths in water poses the question whether light-driven photoconversion is relevant in the mesophotic zone, or whether a different mechanism is involved in the post-translational pigment modification in vivo...
July 4, 2017: International Journal of Molecular Sciences
Mookkandi Palsamy Kesavan, Gujuluva Gangatharan Vinoth Kumar, Kandasamy Anitha, Lokesh Ravi, Jeyaraj Dhaveethu Raja, Gurusamy Rajagopal, Jegathalaprathaban Rajesh
Effective interaction of natural alkaloid Luotonin A (L) and its affixed acceptor molecules 1 and 2 with donor molecule as Bovine serum albumin (BSA) at various pH (4.0, 7.4 and 10.0) medium have been demonstrated using various conventional spectroscopic techniques. These analyses provide some valuable features on the interaction between BSA and acceptor molecules (L, 1 and 2). From the absorption and fluorescence spectral titration studies, the formation of ground-state complexes between the acceptor molecules (L, 1 and 2) and the BSA have been confirmed...
June 27, 2017: Journal of Photochemistry and Photobiology. B, Biology
David M Rees, Oliver Willhoft, Chia-Liang Lin, Rohan Bythell-Douglas, Dale B Wigley
We have developed a novel system to facilitate the rapid and easy cloning of multiple genes (>10) in under a week. Using this system we have been able to successfully clone, overexpress, and purify a number of large multimeric proteins from insect cells, including the chromatin remodeling complexes SWR1 and INO80. Using Förster resonance energy transfer (FRET)-based assays we have demonstrated that our overexpressed enzymes have activities comparable to those purified from sources where the proteins are expressed under their endogenous promoters...
2017: Methods in Enzymology
Chang-Bo Huang, Lin Xu, Jun-Long Zhu, Yu-Xuan Wang, Bin Sun, Xiaopeng Li, Hai-Bo Yang
It is quite challenging to investigate the dynamics of coordination-driven self-assembly due to the existence of multiple intermediates and many possible processes. By taking advantage of the high sensitivity and efficiency of fluorescence-resonance energy transfer (FRET), FRET was successfully employed to real-time monitor the dynamic behavior of coordination-driven self-assembly. The Förster energy transfer efficiencies and kinetic aspects of a series of discrete, well-defined metallacycles have been determined...
July 5, 2017: Journal of the American Chemical Society
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