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Forster resonance energy transfer

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https://www.readbyqxmd.com/read/27930744/interactions-of-calcium-fluctuations-during-cardiomyocyte-contraction-with-real-time-camp-dynamics-detected-by-fret
#1
Julia U Sprenger, Nadja I Bork, Jonas Herting, Thomas H Fischer, Viacheslav O Nikolaev
Calcium (Ca2+) and 3',5'-cyclic adenosine monophosphate (cAMP) play a critical role for cardiac excitation-contraction-coupling. Both second messengers are known to interact with each other, for example via Ca2+-dependent modulation of phosphodiesterase 1 (PDE1) and adenylyl cyclase 5/6 (AC 5/6) activities, which is supposed to occur especially at the local level in distinct subcellular microdomains. Currently, many studies analyze global and local cAMP signaling and its regulation in resting cardiomyocytes devoid of electrical stimulation...
2016: PloS One
https://www.readbyqxmd.com/read/27929144/intermolecular-interactions-in-the-tmem16a-dimer-controlling-channel-activity
#2
Paolo Scudieri, Ilaria Musante, Ambra Gianotti, Oscar Moran, Luis J V Galietta
TMEM16A and TMEM16B are plasma membrane proteins with Ca(2+)-dependent Cl(-) channel function. By replacing the carboxy-terminus of TMEM16A with the equivalent region of TMEM16B, we obtained channels with potentiation of channel activity. Progressive shortening of the chimeric region restricted the "activating domain" to a short sequence close to the last transmembrane domain and led to TMEM16A channels with high activity at very low intracellular Ca(2+) concentrations. To elucidate the molecular mechanism underlying this effect, we carried out experiments based on double chimeras, Forster resonance energy transfer, and intermolecular cross-linking...
December 8, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27928132/improved-fret-biosensor-for-the-measurement-of-bcr-abl-activity-in-chronic-myeloid-leukemia-cells
#3
Mika Horiguchi, Mari Fujioka, Takeshi Kondo, Yoichiro Fujioka, Xinxin Li, Kosui Horiuchi, Aya O Satoh, Prabha Nepal, Shinya Nishide, Asuka Nanbo, Takanori Teshima, Yusuke Ohba
Although the co-development of companion diagnostics with molecular targeted drugs is desirable, truly efficient diagnostics are limited to diseases in which chromosomal translocations or overt mutations are clearly correlated with drug efficacy. Moreover, even for such diseases, few methods are available to predict whether drug administration is effective for each individual patient whose disease is expected to respond to the drug(s). We have previously developed a biosensor based on the principle of Förster resonance energy transfer (FRET) to measure the activity of the tyrosine kinase BCR-ABL and its response to drug treatment in patient-derived chronic myeloid leukemia cells...
December 8, 2016: Cell Structure and Function
https://www.readbyqxmd.com/read/27927983/a-new-method-to-study-heterodimerization-of-membrane-proteins-and-its-application-to-fibroblast-growth-factor-receptors
#4
Nuala Del Piccolo, Sarvenaz Sarabipour, Kalina Hristova
The activity of receptor tyrosine kinases (RTKs) is controlled through their lateral dimerization in the plasma membrane. RTKs are believed to form both homodimers and heterodimers, and the different dimers are believed to play unique roles in cell signaling. However, RTK heterodimers remain poorly characterized, as compared to homodimers, due to limitations in current experimental methods. Here, we develop a Förster Resonance Energy Transfer (FRET)-based methodology to assess the thermodynamics of hetero-interactions in the plasma membrane...
December 7, 2016: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/27924490/site-specific-fluorescent-labeling-of-argonaute-for-fret-based-bio-assays
#5
Sarah Willkomm, Adrian Zander, Dina Grohmann
Deciphering the molecular mechanisms of eukaryotic Argonaute proteins is crucial for the understanding of RNA interference (RNAi), a posttranscriptional gene silencing process. Fluorescence-based single-molecule studies like single-molecule Förster resonance energy transfer (FRET) between a donor and acceptor dye represent a versatile tool to gain a mechanistic understanding of the structural dynamics of a biomolecular complex. Until today it was not possible to site-specifically introduce fluorophores into eukaryotic Argonaute...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27920157/two-serk-receptor-like-kinases-interact-with-the-receptor-like-kinase-ems1-in-anther-cell-fate-determination
#6
Zhiyong Li, Yao Wang, Jian Huang, Nagib Ahsan, Gabriel Biener, Joel Paprocki, Jay J Thelen, Valerica Raicu, Dazhong Zhao
Cell signaling pathways mediated by leucine-rich repeat receptor-like kinases (LRR-RLK) are essential for plant growth, development and defense. The EMS1 (EXCESS MICROSPOROCYTES1) LRR-RLK and its small protein ligand TPD1 (TAPETUM DETERMINANT1) play a fundamental role in somatic and reproductive cell differentiation during early anther development in Arabidopsis (Arabidopsis thaliana). However, it is unclear whether other cell surface molecules serve as co-regulators of EMS1. Here, we show that SERK1 (SOMATIC EMBRYOGENESIS RECEPTOR-LIKE KINASE1) and SERK2 LRR-RLKs act redundantly as co-regulatory and physical partners of EMS1...
December 5, 2016: Plant Physiology
https://www.readbyqxmd.com/read/27916881/optimization-of-cyanine-dye-stability-and-analysis-of-fret-interaction-on-dna-microarrays
#7
Marcel von der Haar, Christopher Heuer, Martin Pähler, Kathrin von der Haar, Patrick Lindner, Thomas Scheper, Frank Stahl
The application of DNA microarrays for high throughput analysis of genetic regulation is often limited by the fluorophores used as markers. The implementation of multi-scan techniques is limited by the fluorophores' susceptibility to photobleaching when exposed to the scanner laser light. This paper presents combined mechanical and chemical strategies which enhance the photostability of cyanine 3 and cyanine 5 as part of solid state DNA microarrays. These strategies are based on scanning the microarrays while the hybridized DNA is still in an aqueous solution with the presence of a reductive/oxidative system (ROXS)...
November 30, 2016: Biology
https://www.readbyqxmd.com/read/27914369/dna-tetrahedral-scaffolds-based-platform-for-the-construction-of-electrochemiluminescence-biosensor
#8
Qiu-Mei Feng, Zhen Zhou, Mei-Xing Li, Wei Zhao, Jing-Juan Xu, Hong-Yuan Chen
Proximal metallic nanoparticles (NPs) could quench the electrochemiluminescence (ECL) emission of semiconductor quantum dots (QDs) due to Förster energy transfer (FRET), but at a certain distance, the coupling of light-emission with surface plasmon resonance (SPR) result in enhanced ECL. Thus, the modification strategies and distances control between QDs and metallic NPs are critical for the ECL intensity of QDs. In this strategy, a SPR enhanced ECL sensor based on DNA tetrahedral scaffolds modified platform was reported for the detection of telomerase activity...
November 27, 2016: Biosensors & Bioelectronics
https://www.readbyqxmd.com/read/27911813/spatiotemporal-imaging-of-small-gtpases-activity-in-live-cells
#9
Stephanie Voss, Dennis M Krüger, Oliver Koch, Yao-Wen Wu
Ras-like small GTPases function as molecular switches and regulate diverse cellular events. To examine the dynamics of signaling requires spatiotemporal visualization of their activity in the cell. Current small GTPase sensors rely on specific effector domains that are available for only a small number of GTPases and compete for endogenous regulator/effector binding. Here, we describe versatile conformational sensors for GTPase activity (COSGAs) based on the conserved GTPase fold. Conformational changes upon GDP/GTP exchange were directly observed in solution, on beads, and in live cells by Förster resonance energy transfer (FRET)...
November 29, 2016: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/27911791/a-functional-role-for-intrinsic-disorder-in-the-tau-tubulin-complex
#10
Ana M Melo, Juliana Coraor, Garrett Alpha-Cobb, Shana Elbaum-Garfinkle, Abhinav Nath, Elizabeth Rhoades
Tau is an intrinsically disordered protein with an important role in maintaining the dynamic instability of neuronal microtubules. Despite intensive study, a detailed understanding of the functional mechanism of tau is lacking. Here, we address this deficiency by using intramolecular single-molecule Förster Resonance Energy Transfer (smFRET) to characterize the conformational ensemble of tau bound to soluble tubulin heterodimers. Tau adopts an open conformation on binding tubulin, in which the long-range contacts between both termini and the microtubule binding region that characterize its compact solution structure are diminished...
November 23, 2016: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/27910894/calcium-ions-function-as-a-booster-of-chromosome-condensation
#11
Rinyaporn Phengchat, Hideaki Takata, Kenichi Morii, Noriko Inada, Hideji Murakoshi, Susumu Uchiyama, Kiichi Fukui
Chromosome condensation is essential for the faithful transmission of genetic information to daughter cells during cell division. The depletion of chromosome scaffold proteins does not prevent chromosome condensation despite structural defects. This suggests that other factors contribute to condensation. Here we investigated the contribution of divalent cations, particularly Ca(2+), to chromosome condensation in vitro and in vivo. Ca(2+) depletion caused defects in proper mitotic progression, particularly in chromosome condensation after the breakdown of the nuclear envelope...
December 2, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27905300/intravital-imaging-of-mouse-urothelium-reveals-activation-of-extracellular-signal-regulated-kinase-by-stretch-induced-intravesical-release-of-atp
#12
Takeshi Sano, Takashi Kobayashi, Hiromitsu Negoro, Atsushi Sengiku, Takuya Hiratsuka, Yuji Kamioka, Louis S Liou, Osamu Ogawa, Michiyuki Matsuda
To better understand the roles played by signaling molecules in the bladder, we established a protocol of intravital imaging of the bladder of mice expressing a Förster/fluorescence resonance energy transfer (FRET) biosensor for extracellular signal-regulated kinase (ERK), which plays critical roles not only in cell growth but also stress responses. With an upright two-photon excitation microscope and a vacuum-stabilized imaging window, cellular ERK activity was visualized in the whole bladder wall, from adventitia to urothelium...
November 2016: Physiological Reports
https://www.readbyqxmd.com/read/27886235/time-lapse-3-d-measurements-of-a-glucose-biosensor-in-multicellular-spheroids-by-light-sheet-fluorescence-microscopy-in-commercial-96-well-plates
#13
Vincent Maioli, George Chennell, Hugh Sparks, Tobia Lana, Sunil Kumar, David Carling, Alessandro Sardini, Chris Dunsby
Light sheet fluorescence microscopy has previously been demonstrated on a commercially available inverted fluorescence microscope frame using the method of oblique plane microscopy (OPM). In this paper, OPM is adapted to allow time-lapse 3-D imaging of 3-D biological cultures in commercially available glass-bottomed 96-well plates using a stage-scanning OPM approach (ssOPM). Time-lapse 3-D imaging of multicellular spheroids expressing a glucose Förster resonance energy transfer (FRET) biosensor is demonstrated in 16 fields of view with image acquisition at 10 minute intervals...
November 25, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27885213/a-fret-biosensor-for-rock-based-on-a-consensus-substrate-sequence-identified-by-kiss-technology
#14
Chunjie Li, Ayako Imanishi, Naoki Komatsu, Kenta Terai, Mutsuki Amano, Kozo Kaibuchi, Michiyuki Matsuda
Genetically-encoded biosensors based on Förster/fluorescence resonance energy transfer (FRET) are versatile tools for studying the spatio-temporal regulation of signaling molecules within not only the cells but also tissues. Perhaps the hardest task in the development of a FRET biosensor for protein kinases is to identify the kinase-specific substrate peptide to be used in the FRET biosensor. To solve this problem, we took advantage of kinase-interacting substrate screening (KISS) technology, which deduces a consensus substrate sequence for the protein kinase of interest...
November 23, 2016: Cell Structure and Function
https://www.readbyqxmd.com/read/27881485/visualization-of-ligand-induced-gi-protein-activation-in-chemotaxing-cells
#15
Kazuyuki Masuda, Jun-Ichi Kitakami, Tohru Kozasa, Tatsuhiko Kodama, Sigeo Ihara, Takao Hamakubo
Cell migration to chemoattractants is critically important in both normal physiology and the pathogenesis of many diseases. In GPCR-mediated chemotaxis, GPCRs transduce the gradient of an extracellular chemotactic ligand into intracellular responses via the activation of heterotrimeric G proteins. However, ligand-induced G protein activation has not been directly imaged as yet in mammalian chemotaxing cells. We developed a Förster resonance energy transfer (FRET) probe (R10-Gi) by linking the Gi protein α subunit to the regulator of G protein signaling domain...
November 23, 2016: FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology
https://www.readbyqxmd.com/read/27873430/synthesis-of-site-specific-dye-labeled-polymer-via-atom-transfer-radical-polymerization-atrp-for-quantitative-characterization-of-the-well-defined-interchain-distance
#16
Ye Sha, Dongliang Qi, Shaochuan Luo, Xinghua Sun, Xiaoliang Wang, Gi Xue, Dongshan Zhou
Novel difunctional initiators that incorporate Förster/fluorescence resonance energy transfer (FRET) pairs are generated to carry out atom transfer radical polymerization of styrene, methyl methacrylate, and n-butyl methacrylate monomers by an efficient manner. Based on the chemical structures of the initiators, the locations of the fluorophore moiety are dictated to be in the center of the chain with accurately quantified chain functionality (>90% labeling ratio). The site-specific integration of FRET dyes into separate polymer chain centers allows for characterization of the well-defined interchain distance quantitatively based on the response between these fluorescent probes...
November 22, 2016: Macromolecular Rapid Communications
https://www.readbyqxmd.com/read/27872188/a-monoclonal-antibody-to-cryptococcus-neoformans-glucuronoxylomannan-manifests-hydrolytic-activity-for-both-peptides-and-polysaccharides
#17
Anthony Bowen, Maggie P Wear, Radames Cordero, Stefan Oscarson, Arturo Casadevall
Studies in the 1980s first showed that some natural antibodies were "catalytic" and able to hydrolyze peptide or phosphodiester bonds in antigens. Many naturally occurring catalytic antibodies have since been isolated from human sera and associated with positive and negative outcomes in autoimmune disease and infection. The function and prevalence of these antibodies, however, remains unclear. A previous study suggested that the 18B7 monoclonal antibody against glucuronoxylomannan (GXM)--the major component of the Cryptococcus neoformans polysaccharide capsule--hydrolyzed a peptide antigen mimetic...
November 21, 2016: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/27871615/a-homogeneous-assay-for-highly-sensitive-detection-of-camv35s-promoter-in-transgenic-soybean-by-f%C3%A3-rster-resonance-energy-transfer-between-nitrogen-doped-graphene-quantum-dots-and-ag-nanoparticles
#18
Yaqi Li, Li Sun, Jing Qian, Chengke Wang, Qian Liu, En Han, Nan Hao, Liuping Zhang, Jianrong Cai, Kun Wang
In this work, a novel homogeneous assay for DNA quantitative analysis based on förster resonance energy transfer (FRET) was developed for cauliflwer mosaic virus 35s (CaMV35S) promoter of transgenic soybean detection. The homogenous FRET of fluorescence signal was fabricated by DNA hybridization with probe modified nitrogen-doped graphene quantum dots (NGQDs) and silver nanoparticles (AgNPs), which acted the donor-acceptor pairs for the first time. The highly efficient FRET and unique properties of the NGQDs made the proposed FRET system as a functionalized detection platform for labelling of DNA...
December 15, 2016: Analytica Chimica Acta
https://www.readbyqxmd.com/read/27869816/mscarlet-a-bright-monomeric-red-fluorescent-protein-for-cellular-imaging
#19
Daphne S Bindels, Lindsay Haarbosch, Laura van Weeren, Marten Postma, Katrin E Wiese, Marieke Mastop, Sylvain Aumonier, Guillaume Gotthard, Antoine Royant, Mark A Hink, Theodorus W J Gadella
We report the engineering of mScarlet, a truly monomeric red fluorescent protein with record brightness, quantum yield (70%) and fluorescence lifetime (3.9 ns). We developed mScarlet starting with a consensus synthetic template and using improved spectroscopic screening techniques; mScarlet's crystal structure reveals a planar and rigidified chromophore. mScarlet outperforms existing red Förster proteins as a fusion tag, and it is especially useful as a fluorescence resonance energy transfer (FRET) acceptor in ratiometric imaging...
November 21, 2016: Nature Methods
https://www.readbyqxmd.com/read/27864764/fret-flim-for-visualizing-and-quantifying-protein-interactions-in-live-plant-cells
#20
Alejandra Freire Rios, Tatyana Radoeva, Bert De Rybel, Dolf Weijers, Jan Willem Borst
Proteins are the workhorses that control most biological processes in living cells. Although proteins can accomplish their functions independently, the vast majority of functions require proteins to interact with other proteins or biomacromolecules. Protein interactions can be investigated through biochemical assays such as co-immunoprecipitation (co-IP) or Western blot analysis, but such assays lack spatial information. Here we describe a well-developed imaging method, Förster resonance energy transfer (FRET) analyzed by fluorescence lifetime imaging microscopy (FLIM), that can be used to visualize protein interactions with both spatial and temporal resolution in live cells...
2017: Methods in Molecular Biology
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