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https://www.readbyqxmd.com/read/28545052/brl3-and-atrgs1-cooperate-to-fine-tune-growth-inhibition-and-ros-activation
#1
Meral Tunc-Ozdemir, Alan M Jones
Plasma membrane-localized leucine-rich repeat receptor-like kinases directly activates G protein complex via interaction with seven transmembrane domain Regulator of G-protein Signaling 1 (AtRGS1) protein. Brassinosteroid insensitive 1 (BRI1) LIKE3 (BRL3) phosphorylates AtRGS1 in vitro. FRET analysis showed that BRL3 and AtRGS1 interaction dynamics change in response to glucose and flg22. Both BRL3 and AtRGS1 function in glucose sensing and brl3 and rgs1-2 single mutants are hyposensitive to high glucose as well as the brl3/rgs1 double mutant...
2017: PloS One
https://www.readbyqxmd.com/read/28544957/zebrafish-as-a-visual-and-dynamic-model-to-study-the-transport-of-nanosized-drug-delivery-systems-across-the-biological-barriers
#2
Ye Li, Xiaoqing Miao, Tongkai Chen, Xiang Yi, Ruibing Wang, Haitao Zhao, Simon Ming-Yuen Lee, Xueqing Wang, Ying Zheng
With the wide application of nanotechnology to drug delivery systems, a simple, dynamic and visual in vivo model for high-throughput screening of novel formulations with fluorescence markers across biological barriers is desperately needed. In vitro cell culture models have been widely used, although they are far from a complimentary in vivo system. Mammalian animal models are common predictive models to study transport, but they are costly and time consuming. Zebrafish (Danio rerio), a small vertebrate model, have the potential to be developed as an "intermediate" model for quick evaluations...
May 10, 2017: Colloids and Surfaces. B, Biointerfaces
https://www.readbyqxmd.com/read/28542243/covalent-dye-attachment-influences-the-dynamics-and-conformational-properties-of-flexible-peptides
#3
Manuel P Luitz, Anders Barth, Alvaro H Crevenna, Rainer Bomblies, Don C Lamb, Martin Zacharias
Fluorescence spectroscopy techniques like Förster resonance energy transfer (FRET) and fluorescence correlation spectroscopy (FCS) have become important tools for the in vitro and in vivo investigation of conformational dynamics in biomolecules. These methods rely on the distance-dependent quenching of the fluorescence signal of a donor fluorophore either by a fluorescent acceptor fluorophore (FRET) or a non-fluorescent quencher, as used in FCS with photoinduced electron transfer (PET). The attachment of fluorophores to the molecule of interest can potentially alter the molecular properties and may affect the relevant conformational states and dynamics especially of flexible biomolecules like intrinsically disordered proteins (IDP)...
2017: PloS One
https://www.readbyqxmd.com/read/28542211/f%C3%A3-rster-resonance-energy-transfer-role-of-diffusion-of-fluorophore-orientation-and-separation-in-observed-shifts-of-fret-efficiency
#4
Bram Wallace, Paul J Atzberger
Förster resonance energy transfer (FRET) is a widely used single-molecule technique for measuring nanoscale distances from changes in the non-radiative transfer of energy between donor and acceptor fluorophores. For macromolecules and complexes this observed transfer efficiency is used to infer changes in molecular conformation under differing experimental conditions. However, sometimes shifts are observed in the FRET efficiency even when there is strong experimental evidence that the molecular conformational state is unchanged...
2017: PloS One
https://www.readbyqxmd.com/read/28540892/aldosterone-synthase-gene-is-not-a-major-susceptibility-gene-for-progression-of-chronic-kidney-disease-in-patients-with-autosomal-dominant-polycystic-kidney-disease
#5
Gnanasambandan Ramanathan, Ramprasad Elumalai, Soundararajan Periyasamy, Bhaskar V K S Lakkakula
Autosomal dominant polycystic kidney disease (ADPKD) is the most common heritable kidney disease and is characterized by bilateral renal cysts. Hypertension is a frequent cause of chronic kidney disease (CKD) and mortality in patients with ADPKD. The aldosterone synthase gene polymorphisms of the renin-angiotensin-aldosterone system have been extensively studied as hypertension candidate genes. The present study is aimed to investigate the potential modifier effect of CYP11B2 gene on the progression of CKD in ADPKD...
May 2017: Saudi Journal of Kidney Diseases and Transplantation
https://www.readbyqxmd.com/read/28539582/fluorescent-rna-cytosine-analogue-an-internal-probe-for-detailed-structure-and-dynamics-investigations
#6
Anders Foller Füchtbauer, Søren Preus, Karl Börjesson, Scott A McPhee, David M J Lilley, L Marcus Wilhelmsson
The bright fluorescent cytosine analogue tC(O) stands out among fluorescent bases due to its virtually unquenched fluorescence emission in duplex DNA. However, like most reported base analogues, it has not been thoroughly characterized in RNA. We here report on the first synthesis and RNA-incorporation of tC(O), and characterize its base-mimicking and fluorescence properties in RNA. As in DNA, we find a high quantum yield inside RNA duplexes (<ΦF> = 0.22) that is virtually unaffected by the neighbouring bases (ΦF = 0...
May 24, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28539409/spatial-analysis-of-cdc42-activity-reveals-a-role-for-plasma-membrane-associated-cdc42-in-centrosome-regulation
#7
Kari A Herrington, Andrew L Trinh, Carolyn Dang, Ellen O'Shaughnessy, Klaus M Hahn, Enrico Gratton, Michelle A Digman, Christine Sütterlin
The ability of the small GTPase Cdc42 to regulate diverse cellular processes depends on tight spatial control of its activity. Cdc42 function is best understood at the plasma membrane (PM), where it regulates cytoskeletal organization and cell polarization. Active Cdc42 has also been detected at the Golgi, but its role and regulation at this organelle is only partially understood. Here we analyze the spatial distribution of Cdc42 activity by monitoring the dynamics of the Cdc42 FLARE biosensor using the phasor approach to FLIM-FRET...
May 24, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28531180/characterization-of-tauc3-antibody-and-demonstration-of-its-potential-to-block-tau-propagation
#8
Samantha B Nicholls, Sarah L DeVos, Caitlin Commins, Chloe Nobuhara, Rachel E Bennett, Diana L Corjuc, Eduardo Maury, Bahareh Eftekharzadeh, Ololade Akingbade, Zhanyun Fan, Allyson D Roe, Shuko Takeda, Susanne Wegmann, Bradley T Hyman
The spread of neurofibrillary tangle (NFT) pathology through the human brain is a hallmark of Alzheimer's disease (AD), which is thought to be caused by the propagation of "seeding" competent soluble misfolded tau. "TauC3", a C-terminally truncated form of tau that is generated by caspase-3 cleavage at D421, has previously been observed in NFTs and has been implicated in tau toxicity. Here we show that TauC3 is found in the seeding competent high molecular weight (HMW) protein fraction of human AD brain. Using a specific TauC3 antibody, we were able to substantially block the HMW tau seeding activity of human AD brain extracts in an in vitro tau seeding FRET assay...
2017: PloS One
https://www.readbyqxmd.com/read/28530838/an-innovative-high-throughput-screening-approach-for-discovery-of-small-molecules-that-inhibit-tnf-receptors
#9
Chih Hung Lo, Nagamani Vunnam, Andrew K Lewis, Ting-Lan Chiu, Benjamin E Brummel, Tory M Schaaf, Benjamin D Grant, Prachi Bawaskar, David D Thomas, Jonathan N Sachs
Tumor necrosis factor receptor 1 (TNFR1) is a transmembrane receptor that binds tumor necrosis factor or lymphotoxin-alpha and plays a critical role in regulating the inflammatory response. Upregulation of these ligands is associated with inflammatory and autoimmune diseases. Current treatments reduce symptoms by sequestering free ligands, but this can cause adverse side effects by unintentionally inhibiting ligand binding to off-target receptors. Hence, there is a need for new small molecules that specifically target the receptors, rather than the ligands...
May 1, 2017: SLAS Discovery
https://www.readbyqxmd.com/read/28529973/high-affinity-mannotetraose-as-an-alternative-to-dextran-in-cona-based-fluorescent-affinity-glucose-assay-due-to-improved-fret-efficiency
#10
Andrea K Locke, Brian M Cummins, Gerard L Coté
Diabetes mellitus affects millions of people worldwide and requires that individuals tightly self-regulate their blood glucose levels to minimize the associated secondary complications. Continuous monitoring devices potentially offer patients a long-term means to tightly monitor their glucose levels. In recent years, fluorescent affinity sensors based on lectins (e.g., Concanavalin A (ConA)) have been implemented in such devices. Traditionally, these sensors pair the lectin with a multivalent ligand, like dextran, in order to develop a competitive binding assay that changes its fluorescent properties in response to the surrounding glucose concentrations...
May 27, 2016: ACS Sensors
https://www.readbyqxmd.com/read/28529063/methodologies-for-studying-the-spliceosome-s-rna-dynamics-with-single-molecule-fret
#11
REVIEW
Clarisse van der Feltz, Aaron A Hoskins
The spliceosome is an extraordinarily dynamic molecular machine in which significant changes in composition as well as protein and RNA conformation are required for carrying out pre-mRNA splicing. Single-molecule fluorescence resonance energy transfer (smFRET) can be used to elucidate these dynamics both in well-characterized model systems and in entire spliceosomes. These types of single-molecule data provide novel information about spliceosome components and can be used to identify sub-populations of molecules with unique behaviors...
May 18, 2017: Methods: a Companion to Methods in Enzymology
https://www.readbyqxmd.com/read/28528543/plasmon-enhanced-energy-transfer-in-photosensitive-nanocrystal-device
#12
Shahab Akhavan, Mehmet Zafer Akgul, Pedro Ludwig Hernandez Martinez, Hilmi Volkan Demir
Förster resonance energy transfer (FRET) interacted with localized surface plasmon (LSP) gives us the ability to overcome inadequate transfer of energy between donor and acceptor nanocrystals (NCs). In this paper, we show LSP-enhanced FRET in colloidal photosensors of NCs in operation, resulting in substantially enhanced photosensitivity. The proposed photosensitive device is a layered self-assembled colloidal platform consisting of separated monolayers of the donor and the acceptor colloidal NCs with an intermediate metal nanoparticle (MNP) layer made of gold inter-spaced by polyelectrolyte layers...
May 20, 2017: ACS Nano
https://www.readbyqxmd.com/read/28525850/highly-sensitive-aptasensor-based-on-fluorescence-resonance-energy-transfer-between-l-cysteine-capped-zns-quantum-dots-and-graphene-oxide-sheets-for-the-determination-of-edifenphos-fungicide
#13
Majid Arvand, Aazam A Mirroshandel
With the advantages of excellent optical properties and biocompatibility, single-strand DNA-functionalized quantum dots have been widely applied in biosensing and bioimaging. A new aptasensor with easy operation, high sensitivity, and high selectivity was developed by immobilizing the aptamer on water soluble l-cysteine capped ZnS quantum dots (QDs). Graphene oxide (GO) sheets are mixed with the aptamer-QDs. Consequently, the aptamer-conjugated QDs bind to the GO sheets to form a GO/aptamer-QDs ensemble. This aptasensor enables the energy transfer based on a fluorescence resonance energy transfer (FRET) from the QDs to the GO sheets, quenching the fluorescence of QDs...
May 12, 2017: Biosensors & Bioelectronics
https://www.readbyqxmd.com/read/28525848/an-amplified-comparative-fluorescence-resonance-energy-transfer-immunosensing-of-ca125-tumor-marker-and-ovarian-cancer-cells-using-green-and-economic-carbon-dots-for-bio-applications-in-labeling-imaging-and-sensing
#14
Somayeh Hamd-Ghadareh, Abdollah Salimi, Fardin Fathi, Saman Bahrami
CA125, is a marker in the clinical diagnosis of several cancers and currently is the best serum-based tumor marker for ovarian cancer. Here, we developed an ultrasensitive antibody-ssDNA aptamer sandwich-type fluorescence immunosensor for CA125 detection. Based on a novel signal amplification strategy the carbon dots (CDs) functionalized with aptamer (CD-aptamer) used as detection probe and PAMAM-Dendrimers/AuNPs was used for covalent attachment of CA125-antibody and completing the sandwich assay method. By measuring of fluorescence resonance energy transfer (FRET) signals between CDs and AuNPs as nanoquenchers, the fluorescence signal quenched during sandwich complex formed between anti-CA125, CA125 and CDs-Aptamer and decreasing of fluorescence response signal is related to CA125 concentrations...
May 4, 2017: Biosensors & Bioelectronics
https://www.readbyqxmd.com/read/28524818/fluorescence-lifetime-imaging-microscopy-reveals-rerouting-of-snare-trafficking-driving-dendritic-cell-activation
#15
Daniëlle Rianne José Verboogen, Natalia González Mancha, Martin Ter Beest, Geert van den Bogaart
SNARE proteins play a crucial role in intracellular trafficking by catalyzing membrane fusion, but assigning SNAREs to specific intracellular transport routes is challenging with current techniques. We developed a novel Förster resonance energy transfer-fluorescence lifetime imaging microscopy (FRET-FLIM)-based technique allowing visualization of real-time local interactions of fluorescently tagged SNARE proteins in live cells. We used FRET-FLIM to delineate the trafficking steps underlying the release of the inflammatory cytokine interleukin-6 (IL-6) from human blood-derived dendritic cells...
May 19, 2017: ELife
https://www.readbyqxmd.com/read/28524357/double-caging-linker-for-and-type-fluorogenic-construction-of-protein-antibody-bioconjugates-and-in-situ-quantification
#16
Shiyong Liu, Guhuan Liu, Guohai Shi, Haoyue Sheng, Yanyan Jiang, Haojun Liang
We report on in situ fluorescent quantification of the conjugation efficiency between azide-terminated synthetic polymers/ imaging probes and thiol-functionalized antibodies/proteins/peptides, by utilizing a doubly caged profluorescent and heterodifunctional core molecule (C1) as the self-sorting bridging unit. Orthogonal dual 'click' coupling of C1 with azide- and thiol-functionalized precursors leads to highly fluorescent bioconjugates, whereas single click products of C1 remain essentially nonfluorescent...
May 19, 2017: Angewandte Chemie
https://www.readbyqxmd.com/read/28522796/effects-of-%C3%AE-adrenoceptor-and-catechol-o-methyl-transferase-comt-polymorphism-on-postoperative-outcome-in-cardiac-surgery-patients
#17
Stefan Dhein, Pascal M Dohmen, Matthias Sauer, Julia Tews, Johannes Weickmann, Anne-Kathrin Funkat, Martin Misfeld, Michael A Borger, Friedrich W Mohr
BACKGROUND There is a long-standing debate about the role of beta-adrenoceptor polymorphisms in the cardiovascular system. We wanted to elucidate whether β1-adrenoceptor-polymorphisms affects the postoperative catecholamine consumption and the length of intermediate care unit stay in patients undergoing cardiac surgery, and whether this might be enhanced or attenuated by catechol-O-methyl-transferase (COMT) polymorphism. MATERIAL AND METHODS We included 116 patients (63±1.2 years; 34% females; 81±1 kg) undergoing cardiac surgery...
May 19, 2017: Medical Science Monitor Basic Research
https://www.readbyqxmd.com/read/28522396/ultrafast-spectroscopy-on-dna-cleavage-by-endonuclease-in-molecular-crowding
#18
Priya Singh, Susobhan Choudhury, Shreyasi Dutta, Aniruddha Adhikari, Siddhartha Bhattacharya, Debasish Pal, Samir Kumar Pal
The jam-packed intracellular environments differ the activity of a biological macromolecule from that in laboratory environments (in vitro) through a number of mechanisms called molecular crowding related to structure, function and dynamics of the macromolecule. Here, we have explored the structure, function and dynamics of a model enzyme protein DNase I in molecular crowing of polyethylene glycol (PEG; MW 3350). We have used steady state and picosecond resolved dynamics of a well-known intercalator ethidium bromide (EB) in a 20-mer double-stranded DNA (dsDNA) to monitor the DNA-cleavage by the enzyme in absence and presence PEG...
May 15, 2017: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/28520430/fluorescence-dynamics-of-a-fret-probe-designed-for-crowding-studies
#19
Megan Currie, Hannah Leopold, Jacob Schwarz, Arnold J Boersma, Erin D Sheets, Ahmed A Heikal
Living cells are crowded with macromolecules and organelles. As a result, there is an urgent need for molecular sensors for quantitative, site-specific assessment of the macromolecular crowding effects on a myriad of biochemical processes towards quantitative cell biology and biophysics. Here we investigate the excited-state dynamics and translational diffusion of a novel FRET sensor (mCerulean-linker-mCitrine) in a buffer (PBS, pH 7.4) at room temperature. Complementary experiments were carried out on free CFP, YFP, and the cleaved FRET probe as controls...
May 18, 2017: Journal of Physical Chemistry. B
https://www.readbyqxmd.com/read/28516765/peptide-microarray-based-metal-enhanced-fluorescence-assay-for-multiple-profiling-matrix-metalloproteinases-activities
#20
Zhen Lei, Hua Zhang, Yaoqi Wang, Xianying Meng, Zhenxin Wang
Matrix metalloproteinases (MMPs) are closely associated with cancer cell invasion and metastasis. Herein, a fluorescence resonance energy transfer (FRET)-peptide microarray-based metal enhanced fluorescence (MEF) assay is proposed for multiple and sensitive profiling MMPs activities on a novel Au/Ag@SiO2 substrate. The Au/Ag@SiO2 substrate is prepared by electroless deposition of silver on gold nanoparticle (GNP) seeds, followed by SiO2 shell coating and surface functionalization. The specific FRET peptides are spotted on the Au/Ag@SiO2 substrate to sensitively detect MMPs (MMP-2, 3, 7, 9, 14) via fluorescence recovery by the MMP cleavage of quenched peptide motifs, and further enhanced by MEF...
May 18, 2017: Analytical Chemistry
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