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https://www.readbyqxmd.com/read/28338736/microcompartmentation-of-cytosolic-aldolase-by-interaction-with-the-actin-cytoskeleton-in-arabidopsis
#1
Constantine Garagounis, Kalliopi-Ioanna Kostaki, Tim J Hawkins, Ian Cummins, Mark D Fricker, Patrick J Hussey, Alistair M Hetherington, Lee J Sweetlove
Evidence is accumulating for molecular microcompartments formed when proteins interact in localized domains with the cytoskeleton, organelle surfaces, and intracellular membranes. To understand the potential functional significance of protein microcompartmentation in plants, we studied the interaction of the glycolytic enzyme fructose bisphosphate aldolase with actin in Arabidopsis thaliana. Homology modelling of a major cytosolic isozyme of aldolase, FBA8, suggested that the tetrameric holoenzyme has two actin binding sites and could therefore act as an actin-bundling protein, as was reported for animal aldolases...
February 17, 2017: Journal of Experimental Botany
https://www.readbyqxmd.com/read/28338043/redshifted-cherenkov-radiation-for-in-vivo-imaging-coupling-cherenkov-radiation-energy-transfer-to-multiple-f%C3%A3-rster-resonance-energy-transfers
#2
Yann Bernhard, Bertrand Collin, Richard A Decréau
Cherenkov Radiation (CR), this blue glow seen in nuclear reactors, is an optical light originating from energetic β-emitter radionuclides. CR emitter (90)Y triggers a cascade of energy transfers in the presence of a mixed population of fluorophores (which each other match their respective absorption and emission maxima): Cherenkov Radiation Energy Transfer (CRET) first, followed by multiple Förster Resonance Energy transfers (FRET): CRET ratios were calculated to give a rough estimate of the transfer efficiency...
March 24, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28337497/dimerization-and-conformation-related-free-energy-landscapes-of-dye-tagged-amyloid-%C3%AE-12-28-linked-to-fret-experiments
#3
Alexander Kulesza, Steven Daly, Philippe Dugourd
We have investigated the free energy landscape of Aβ-peptide dimer models in connection to gas-phase FRET experiments. We use a FRET-related distance coordinate and one conformation-related coordinate per monomer for accelerated structural exploration with well-tempered metadynamics in solvent and in vacuo. The free energy profiles indicate that FRET under equilibrium conditions should be significantly affected by the de-solvation upon the transfer of ions to the gas-phase. In contrast, a change in the protonation state is found to be less impacting once de-solvated...
March 24, 2017: Physical Chemistry Chemical Physics: PCCP
https://www.readbyqxmd.com/read/28336533/critical-residues-and-motifs-for-homodimerization-of-the-first-transmembrane-domain-of-the-plasma-membrane-glycoprotein-cd36
#4
Peng Wei, Fu-de Sun, Li-Min Zuo, Jing Qu, Peng Chen, Li-da Xu, Shi-Zhong Luo
The plasma transmembrane (TM) glycoprotein CD36 is critically involved in many essential signaling processes, especially the binding /uptake of long-chain fatty acids and oxidized low-density lipoproteins. The association of CD36 potentially activates cytosolic protein tyrosine kinases that are thought to associate with the C-terminal cytoplasmic tail of CD36. To understand the mechanisms by which CD36 mediates ligand binding and signal transduction, we have characterized the homo- oligomeric interaction of CD36 TM domains in membrane environments and with molecular dynamics (MD) simulations...
March 23, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28336075/engineering-bret-sensor-proteins
#5
Remco Arts, Stijn J A Aper, Maarten Merkx
FRET-sensors have become important tools for intracellular imaging, but their dependence on external illumination presents some limitations, such as photobleaching and phototoxicity, which limit measurements over extended periods of time. Fluorescence measurements also suffer from autofluorescence and light scattering, which hampers in vivo imaging and measurements in strongly absorbing and scattering media such as blood. In principle, these issues can be resolved by using sensors based on bioluminescence resonance energy transfer (BRET)...
2017: Methods in Enzymology
https://www.readbyqxmd.com/read/28336060/recent-advances-in-development-of-genetically-encoded-fluorescent-sensors
#6
Lynn Sanford, Amy Palmer
Genetically encoded fluorescent sensors are essential tools in modern biological research, and recent advances in fluorescent proteins (FPs) have expanded the scope of sensor design and implementation. In this review we compare different sensor platforms, including Förster resonance energy transfer (FRET) sensors, fluorescence-modulated single FP-based sensors, translocation sensors, complementation sensors, and dimerization-based sensors. We discuss elements of sensor design and engineering for each platform, including the incorporation of new types of FPs and sensor screening techniques...
2017: Methods in Enzymology
https://www.readbyqxmd.com/read/28334690/intracellular-fluorescent-light-up-bioprobes-with-different-morphology-for-image-guided-photothermal-cancer-therapy
#7
Bangbang Li, Peng Zhang, Jianwei Du, Xiao Zhao, Youxiang Wang
Multifunctional nanoprobe was drawing increased attention in tumor diagnosis and therapy. The simple and effective establishment of the theranostic nanoplatforms was still under urgent need. Meanwhile, the targeting ability and morphology of nanoprobe were essential for the effective endocytosis, which could further affect the diagnosis. In this work, two morphologies of nanoprobes were fabricated using gold nanorods (AuNRs) and gold nanospheres (AuNSs). Thiolated-hyaluronic acid labeled with nile blue (HS-HA-NB), a near-infrared (NIR) fluorescence dye, was coated on the surface of the gold nanoparticles to form stable nanoprobes (AuNR@HS-HA-NB, AuNS@HS-HA-NB)...
March 9, 2017: Colloids and Surfaces. B, Biointerfaces
https://www.readbyqxmd.com/read/28333446/quantification-of-membrane-protein-self-association-with-a-high-throughput-compatible-fluorescence-assay
#8
Junbei Li, Xiaoyan Jade Qiu
For many membrane proteins, self-association serves both structural and functional roles. Studies of such association can be simplified by switching to micelles as the membrane-mimicking environment. But native interaction is not preserved in all detergents. The selection of suitable conditions for biochemical experiments would be greatly facilitated by a quantitative high-throughput assay. Here we showed that the fluorescence polarization (FP) reduction, resulted from homo-FRET and measured in a high-throughput compatible format, can be used to determine both association states and constants for membrane proteins in micelles...
March 23, 2017: Biochemistry
https://www.readbyqxmd.com/read/28332923/different-types-of-pausing-modes-during-transcription-initiation
#9
Eitan Lerner, Antonino Ingargiola, Jookyung J Lee, Sergei Borukhov, Xavier Michalet, Shimon Weiss
In many cases, initiation is rate limiting to transcription. This due in part to the multiple cycles of abortive transcription that delay promoter escape and the transition from initiation to elongation. Pausing of transcription in initiation can further delay promoter escape. The previously hypothesized pausing in initiation was confirmed by two recent studies from Duchi et al. (1) and from Lerner, Chung et al. (2) In both studies, pausing is attributed to lack of forward translocation of the nascent transcript during initiation...
March 23, 2017: Transcription
https://www.readbyqxmd.com/read/28332666/direct-observation-of-breathing-dynamics-at-the-mismatch-induced-dna-bubble-with-nanometre-accuracy-a-smfret-study
#10
Tapas Paul, Subhas C Bera, Padmaja P Mishra
The detailed conformational dynamics of the melted region in double-stranded DNA has been studied using a combination of ensemble and single-molecule FRET techniques. We monitored the millisecond time scale fluctuation kinetics of the two strands at the bubble region that varies with the size of the bubble. As the individual strands at the melting bubble behave as single-stranded DNA, and hence fluctuate dynamically to attain energetically favored configurations, the rates of these fluctuations increase with increase in the bubble size...
March 23, 2017: Nanoscale
https://www.readbyqxmd.com/read/28332566/a-conformational-change-within-the-wave2-complex-regulates-its-degradation-following-cellular-activation
#11
Noah Joseph, Guy Biber, Sophia Fried, Barak Reicher, Omer Levy, Batel Sabag, Elad Noy, Mira Barda-Saad
WASp family Verprolin-homologous protein-2 (WAVE2), a member of the Wiskott-Aldrich syndrome protein (WASp) family of actin nucleation promoting factors, is a central regulator of actin cytoskeleton polymerization and dynamics. Multiple signaling pathways operate via WAVE2 to promote the actin-nucleating activity of the actin-related protein 2/3 (Arp2/3) complex. WAVE2 exists as a part of a pentameric protein complex known as the WAVE regulatory complex (WRC), which is unstable in the absence of its individual proteins...
March 23, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28332150/dynamic-nature-of-presenilin1-%C3%AE-secretase-implication-for-alzheimer-s-disease-pathogenesis
#12
REVIEW
Katarzyna Marta Zoltowska, Oksana Berezovska
Presenilin 1 (PS1) is a catalytic component of the γ-secretase complex, responsible for the intramembraneous cleavage of more than 90 type I transmembrane proteins, including Alzheimer's disease (AD)-related amyloid precursor protein (APP). The γ-secretase-mediated cleavage of the APP C-terminal membrane stub leads to the production of various amyloid β (Aβ) species. The assembly of Aβ into neurotoxic oligomers, which causes synaptic dysfunction and neurodegeneration, is influenced by the relative ratio of the longer (Aβ42/43) to shorter Aβ (Aβ40) peptides...
March 22, 2017: Molecular Neurobiology
https://www.readbyqxmd.com/read/28330769/difference-in-the-core-shell-dynamics-of-polyethyleneimine-and-poly-l-lysine-dna-polyplexes
#13
Elina Vuorimaa-Laukkanen, Ekaterina S Lisitsyna, Tiia-Maaria Ketola, Emmanuelle Morin-Pickardat, Huamin Liang, Martina Hanzlíková, Marjo Yliperttula
Electrostatic polymer-DNA complexes (polyplexes) have been widely investigated for DNA delivery, and remarkable differences in transfection efficacy have been seen among the materials. For example, polyethyleneimine (PEI) mediates DNA transfection more effectively than poly(l-lysine) (PLL). Biophysical properties of the polyplexes may explain their different properties in gene delivery. We investigated the structural dynamics in DNA polyplexes, especially the material exchange between the core and shell regions of the PEI and PLL polyplexes...
March 18, 2017: European Journal of Pharmaceutical Sciences
https://www.readbyqxmd.com/read/28329679/mammalian-diaphanous-1-mediates-a-pathway-for-e-cadherin-to-stabilize-epithelial-barriers-through-junctional-contractility
#14
Bipul R Acharya, Selwin K Wu, Zi Zhao Lieu, Robert G Parton, Stephan W Grill, Alexander D Bershadsky, Guillermo A Gomez, Alpha S Yap
Formins are a diverse class of actin regulators that influence filament dynamics and organization. Several formins have been identified at epithelial adherens junctions, but their functional impact remains incompletely understood. Here, we tested the hypothesis that formins might affect epithelial interactions through junctional contractility. We focused on mDia1, which was recruited to the zonula adherens (ZA) of established Caco-2 monolayers in response to E-cadherin and RhoA. mDia1 was necessary for contractility at the ZA, measured by assays that include a FRET-based sensor that reports molecular-level tension across αE-catenin...
March 21, 2017: Cell Reports
https://www.readbyqxmd.com/read/28328200/fluorescence-resonance-energy-transfer-based-dna-tetrahedron-nano-tweezer-for-highly-reliable-detection-of-tumor-related-mrna-in-living-cells
#15
Lei He, Dan-Qing Lu, Hao Liang, Sitao Xie, Can Luo, Miaomiao Hu, Liujun Xu, Xiaobing Zhang, Weihong Tan
Accurate detection and imaging of tumor-related mRNA in living cells hold great promise for early cancer detection. However, currently, most probes designed to image intracellular mRNA confront intrinsic interferences arising from complex biological matrices and resulting in inevitable false-positive signals. To circumvent this problem, a new class of intracellular DNA nanoprobe, termed DNA tetrahedron nano-tweezer (DTNT), was developed to reliably image tumor-related mRNA in living cells based on the FRET (fluorescence resonance energy transfer) "off" to "on" signal readout mode...
March 22, 2017: ACS Nano
https://www.readbyqxmd.com/read/28327735/reverse-micelle-based-water-soluble-nanoparticles-for-simultaneous-bioimaging-and-drug-delivery
#16
Ying Chen, Yong Liu, Yongchao Yao, Shiyong Zhang, Zhongwei Gu
With special confined water pools, reverse micelles (RMs) have shown potential for a wide range of applications. However, the inherent water-insolubility of RMs hinders their further application prospects, especially for applications related to biology. We recently reported the first successful transfer of RMs from organic media to an aqueous phase without changing the smart water pools by the hydrolysis of an arm-cleavable interfacial cross-linked reverse micelles. Herein, we employed another elaborate amphiphile 1 to construct new acrylamide-based cross-linked water-soluble nanoparticles (ACW-NPs) under much gentler conditions...
March 22, 2017: Organic & Biomolecular Chemistry
https://www.readbyqxmd.com/read/28327558/actomyosin-generated-tension-on-cadherin-is-similar-between-dividing-and-non-dividing-epithelial-cells-in-early-xenopus-laevis-embryos
#17
Gaëtan Herbomel, Guillaume Hatte, Julien Roul, Sergi Padilla-Parra, Jean-Pierre Tassan, Marc Tramier
Epithelia represent a unique situation where polarized cells must maintain sufficiently strong cell-cell contacts to guarantee the epithelial integrity indispensable for barrier functions. Nevertheless, epithelia must also keep sufficient plasticity which is crucial during development and morphogenesis. Adherens junctions and mechanical forces produced by the actomyosin cytoskeleton are major players for epithelial integrity maintenance and plasticity regulations. To understand how the epithelium is able to meet such a challenge, it is indispensable to determine how cellular junctions and mechanical forces acting at adherens junctions are regulated...
March 22, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28325045/particle-size-dependent-f%C3%A3-rster-resonance-energy-transfer-from-upconversion-nanoparticles-to-organic-dyes
#18
Verena Muhr, Christian Würth, Marco Kraft, Markus Buchner, Ute Resch-Genger, Antje J Baeumner, Thomas Hirsch
Upconversion nanoparticles (UCNPs) are attractive candidates for energy transfer-based analytical applications. In contrast to classical donor-acceptor pairs, these particles contain many emitting lanthanide ions together with numerous acceptor dye molecules at different distances to each other, strongly depending on the particle diameter. UCNPs with precisely controlled sizes between 10 and 43 nm were prepared and functionalized with rose bengal and sulforhodamine B by a ligand exchange procedure. Time-resolved studies of the upconversion luminescence of the UCNP donor revealed a considerable shortening of the donor lifetime as a clear hint for Förster resonance energy transfer (FRET)...
March 22, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28324603/fret-microscopy-for-real-time-visualization-of-second-messengers-in-living-cells
#19
Axel E Kraft, Viacheslav O Nikolaev
Förster Resonance Energy Transfer (FRET) microscopy is a useful tool in molecular biology and medical research to monitor and quantify real-time dynamics of protein-protein interactions and biochemical processes. Using this well-established technique, many novel signaling mechanisms can be investigated in intact cells or tissues and even in various subcellular compartments. Here, we describe how to perform FRET measurements in living cells expressing FRET-based biosensors and how to evaluate these data. This general protocol can be applied for FRET measurements with various fluorescent biosensors...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28317958/the-collective-behavior-of-spring-like-motifs-tethered-to-a-dna-origami-nanostructure
#20
E-C Schöneweiß, B Saccà
Dynamic DNA nanotechnology relies on the integration of small switchable motifs at suitable positions of DNA nanostructures, thus enabling the manipulation of matter with nanometer spatial accuracy in a trigger-dependent fashion. Typical examples of such motifs are hairpins, whose elongation into duplexes can be used to perform long-range, translational movements. In this work, we used temperature-dependent FRET spectroscopy to determine the thermal stabilities of distinct sets of hairpins integrated into the central seam of a DNA origami structure...
March 20, 2017: Nanoscale
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