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https://www.readbyqxmd.com/read/28634884/a-nanosensor-based-on-carbon-dots-for-recovered-fluorescence-detection-clenbuterol-in-pork-samples
#1
Yalan Liu, Qiujun Lu, Xiaojun Hu, Haiyan Wang, Haitao Li, Youyu Zhang, Shouzhuo Yao
Clenbuterol (CLB), a member of β-agonist family, has now been a serious threat to human health due to its illegal usage in the animal feed. In this paper, we designed a fluorescence resonance energy transfer (FRET) system consisting of carbon dots (C-dots) and gold nanoparticles (AuNPs) for recovered fluorescence detecting of CLB. In the presence of CLB, CLB molecules can interact with AuNPs via Au-N bonds, preventing the interaction of C-dots and AuNPs, which induced the recover of the fluorescent intensity...
June 20, 2017: Journal of Fluorescence
https://www.readbyqxmd.com/read/28634298/analyses-of-pde-regulated-phosphoproteomes-reveal-unique-and-specific-camp-signaling-modules-in-t-cells
#2
Michael-Claude G Beltejar, Ho-Tak Lau, Martin G Golkowski, Shao-En Ong, Joseph A Beavo
Specific functions for different cyclic nucleotide phosphodiesterases (PDEs) have not yet been identified in most cell types. Conventional approaches to study PDE function typically rely on measurements of global cAMP, general increases in cAMP-dependent protein kinase (PKA), or the activity of exchange protein activated by cAMP (EPAC). Although newer approaches using subcellularly targeted FRET reporter sensors have helped define more compartmentalized regulation of cAMP, PKA, and EPAC, they have limited ability to link this regulation to downstream effector molecules and biological functions...
June 20, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28633196/tight-junction-strand-formation-by-claudin-10-isoforms-and-claudin-10a-10b-chimeras
#3
Susanne Milatz, Jörg Piontek, Caroline Hempel, Luca Meoli, Christoph Grohe, Anja Fromm, In-Fah M Lee, Rukeia El-Athman, Dorothee Günzel
Claudins are integral components of tight junctions (TJs) in epithelia and endothelia. When expressed in cell lines devoid of TJs, claudins are able to form TJ-like strands at contacts between adjacent cells. According to a current model of TJ strand formation, claudin protomers assemble in an antiparallel double row within the plasma membrane of each cell (cis-interaction) while binding to corresponding double rows from the neighboring cells (trans-interaction). Cis-interaction was proposed to involve two interfaces of the protomers' first extracellular segment (extracellular loop (ECL)1)...
June 20, 2017: Annals of the New York Academy of Sciences
https://www.readbyqxmd.com/read/28631482/carbohydrate-bis-acetal-based-substrates-as-tunable-fluorescence-quenched-probes-for-monitoring-exo-glycosidase-activity
#4
Samy Cecioni, David J Vocadlo
Tunable Förster resonance energy transfer (FRET)-quenched substrates are useful for monitoring the activity of various enzymes within their relevant physiological environments. Development of FRET-quenched substrates for exo-glycosidases, however, has been hindered by their constrained pocket-shaped active sites. Here we report the design of a new class of substrate that overcomes this problem. These Bis-Acetal-Based Substrates (BABS) bear a hemiacetal aglycon leaving group that tethers fluorochromes in close proximity, also positioning them distant from the active site pocket...
June 20, 2017: Journal of the American Chemical Society
https://www.readbyqxmd.com/read/28631479/quantitative-analysis-of-ligand-induced-heterodimerization-of-two-distinct-receptors
#5
Chang Lu, Zhi-Xin Wang
The induced dimerization of two distinct receptors through a heterobifunctional inducer is prevalent among all levels of cellular signaling processes, yet its complexity poses difficulty for systematic quantitative analysis. This paper first shows how to calculate the amount of any possible complex or monomer of heteroligand and two receptors present at equilibrium. The theory is subsequently applied to the determination of three independent equilibrium parameters involved in the rapamycin induced FKBP and FRB dimerization, in which all parameters were simultaneously estimated using one set of fluorescence resonance energy transfer (FRET) experiments...
June 22, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28630044/single-molecule-fret-analysis-discloses-the-dynamics-of-the-dna-topoisomerase-ii-top2-interaction-in-the-presence-of-top2-targeting-agents
#6
Wan-Chen Huang, Chun-Ying Lee, Tao-Shih Hsieh
Topoisomerases play crucial roles in DNA replication, transcription, and recombination. For instance, topoisomerase II (Top2) is critically important for resolving DNA tangles during cell division, and as such, it is a broad anticancer drug target. Top2 regulates DNA topology by transiently breaking one double stranded DNA molecule (cleavage), allowing a second double strand to pass through the opened DNA gate (opening), and then closing the gate by rejoining the broken ends. Drugs that modulate Top2 catalysis may therefore affect enzymatic activity at several different steps...
June 19, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28630006/multiple-conformational-states-of-dnaa-protein-regulate-its-interaction-with-dnaa-boxes-in-the-initiation-of-dna-replication
#7
Meera J Patel, Lavesh Bhatia, Gulden Yilmaz, Esther E Biswas-Fiss, Subhasis B Biswas
DnaA protein is the initiator of genomic DNA replication in prokaryotes. It binds to specific DNA sequences in the origin of DNA replication and unwinds small AT-rich sequences downstream for the assembly of the replisome. The mechanism of activation of DnaA that enables it to bind and organize the origin DNA and leads to replication initiation remains unclear. In this study, we have developed double-labeled fluorescent DnaA probes to analyze conformational states of DnaA protein upon binding DNA, nucleotide, and Soj sporulation protein using Fluorescence Resonance Energy Transfer (FRET)...
June 16, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28629033/a-new-finding-on-the-in-vivo-crevice-corrosion-damage-in-a-cocrmo-hip-implant
#8
Reza H Oskouei, Mohammad Reza Barati, Hamidreza Farhoudi, Mark Taylor, Lucian Bogdan Solomon
A detailed investigation was performed to characterize the fretting wear and corrosion damage to the neck component of a CoCrMo stem from a metal-on-polyethylene implant retrieved after 99months. The stem was a low-carbon (0.07wt%) wrought Co-28Cr-6Mo alloy with no secondary carbide phases in the matrix (γ-phase). The original design of the neck surface contained an intentionally fabricated knurled profile with a valley-to-peak range of approximately 11μm. Roughness measurements indicated that the tip of the knurled profile was significantly damaged, especially in the distal medial region of the neck, with up to a 22% reduction in the mean peak-to-valley height (Ra) compared to the original profile...
October 1, 2017: Materials Science & Engineering. C, Materials for Biological Applications
https://www.readbyqxmd.com/read/28627166/self-assembled-conjugated-polymer-chitosan-graft-oleic-acid-micelles-for-fast-visible-detection-of-aliphatic-biogenic-amines-by-a-turn-on-fret
#9
Haoquan Zhong, Chunchen Liu, Wenjiao Ge, Run-Cang Sun, Fei Huang, Xiaohui Wang
A fluorescent "turn-on" sensor for fast measuring aliphatic biogenic amines (BAs) was developed based on fluorescent carboxylated polyfluorene (PFTBTCOOH)/ chitosan-graft-oleic acid (CS-graft-OA) micelles. In this system, biobased CS-graft-OA micelles prepared by graft modification of chitosan with oleic acid serve as the vector of hydrophobic fluorescent probe. Bright "turn-on" fluorescence was achieved by the intermolecular fluorescence resonance energy transfer (FRET) of the encapsulated PFTBTCOOH driven by the formation of electrostatic complex with aliphatic BAs, which facilitate a visual identification...
June 19, 2017: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/28625737/insights-into-the-conformations-and-dynamics-of-intrinsically-disordered-proteins-using-single-molecule-fluorescence
#10
REVIEW
Gregory-Neal Gomes, Claudiu C Gradinaru
Most proteins are not static structures, but many of them are found in a dynamic state, exchanging conformations on various time scales as a key aspect of their biological function. An entire spectrum of structural disorder exists in proteins and obtaining a satisfactory quantitative description of these states remains a challenge. Single-molecule fluorescence spectroscopy techniques are uniquely suited for this task, by measuring conformations without ensemble averaging and kinetics without interference from asynchronous processes...
June 15, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28625686/the-influence-of-contamination-and-cleaning-on-the-strength-of-modular-head-taper-fixation-in-total-hip-arthroplasty
#11
Annika Krull, Michael M Morlock, Nicholas E Bishop
BACKGROUND: Intraoperative interface contamination of modular head-stem taper junctions of hip implants can lead to poor fixation strength, causing fretting and crevice corrosion or even stem taper fracture. Careful cleaning before assembly should help to reduce these problems. The purpose of this study was to determine the effect of cleaning (with and without drying) contaminated taper interfaces on the taper fixation strength. METHODS: Metal or ceramic heads were impacted onto titanium alloy stem tapers with cleaned or contaminated (fat or saline solution) interfaces...
May 13, 2017: Journal of Arthroplasty
https://www.readbyqxmd.com/read/28615444/the-human-mitochondrial-single-strand-dna-binding-protein-displays-distinct-kinetics-and-thermodynamics-of-dna-binding-and-exchange
#12
Yufeng Qian, Kenneth A Johnson
The human mitochondrial SSB (mtSSB) is a homo-tetrameric protein, involved in mtDNA replication and maintenance. Although mtSSB is structurally similar to SSB from Escherichia coli (EcoSSB), it lacks the C-terminal disordered domain but little is known about the biophysics of mtSSB-ssDNA interactions. Here, we characterized the kinetics and thermodynamics of mtSSB binding to ssDNA by equilibrium titrations and stopped-flow kinetic measurements. We show that the mtSSB tetramer can bind to ssDNA in two distinct binding modes: (SSB)30 and (SSB)60, defined by DNA binding site sizes of 30 and 60 nt, respectively...
June 14, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28613885/towards-complete-sequence-flexibility-of-nucleic-acid-base-analogue-fret
#13
Moa Sandberg Wranne, Anders Foller Füchtbauer, Blaise Dumat, Mattias Sven Risto Bood, Afaf H El-Sagheer, Tom Brown, Henrik Gradén, Morten Grøtli, L Marcus Wilhelmsson
Förster resonance energy transfer (FRET) using fluorescent base analogues is a powerful means of obtaining high-resolution nucleic acid structure and dynamics information that favorably complements techniques such as NMR and X-ray crystal-lography. Here we expand the base-base FRET repertoire with an adenine analogue FRET-pair. Phosphoramidite-protected quadracyclic 2'-deoxyadenosine analogues qAN1 (donor) and qAnitro (acceptor) were synthesized and incorporated into DNA by a generic, reliable, and high-yielding route, and both constitute excellent adenine analogues...
June 14, 2017: Journal of the American Chemical Society
https://www.readbyqxmd.com/read/28612865/esipt-and-fret-probes-for-monitoring-nanoparticle-polymer-coating-stability
#14
Julio C Azcárate, Sebastián A Díaz, Jonathan A Fauerbach, Florencia Gillanders, Aldo A Rubert, Elizabeth A Jares-Erijman, Thomas M Jovin, Mariano H Fonticelli
Coating strategies of inorganic nanoparticles (NPs) can provide properties unavailable to the NP core alone, such as targeting, specific sensing, and increased biocompatibility. Non-covalent amphiphilic NP capping polymers function via hydrophobic interactions with surface ligands and are extensively used to transfer NPs to aqueous media. For applications of coated NPs as actuators (sensors, markers, or for drug delivery) in a complex environment, such as biological systems, it is important to achieve a deep understanding of the factors affecting coating stability and behavior...
June 14, 2017: Nanoscale
https://www.readbyqxmd.com/read/28611216/ubiquitin-s65-phosphorylation-engenders-a-ph-sensitive-conformational-switch
#15
Xu Dong, Zhou Gong, Yun-Bi Lu, Kan Liu, Ling-Yun Qin, Meng-Lin Ran, Chang-Li Zhang, Zhu Liu, Wei-Ping Zhang, Chun Tang
Ubiquitin (Ub) is an important signaling protein. Recent studies have shown that Ub can be enzymatically phosphorylated at S65, and that the resulting pUb exhibits two conformational states-a relaxed state and a retracted state. However, crystallization efforts have yielded only the structure for the relaxed state, which was found similar to that of unmodified Ub. Here we present the solution structures of pUb in both states obtained through refinement against state-specific NMR restraints. We show that the retracted state differs from the relaxed state by the retraction of the last β-strand and by the extension of the second α-helix...
June 13, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28607089/weak-protein-protein-interactions-in-live-cells-are-quantified-by-cell-volume-modulation
#16
Shahar Sukenik, Pin Ren, Martin Gruebele
Weakly bound protein complexes play a crucial role in metabolic, regulatory, and signaling pathways, due in part to the high tunability of their bound and unbound populations. This tunability makes weak binding (micromolar to millimolar dissociation constants) difficult to quantify under biologically relevant conditions. Here, we use rapid perturbation of cell volume to modulate the concentration of weakly bound protein complexes, allowing us to detect their dissociation constant and stoichiometry directly inside the cell...
June 12, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28607002/atm-wip1-activities-at-chromatin-control-plk1-re-activation-to-determine-g2-checkpoint-duration
#17
Himjyot Jaiswal, Jan Benada, Erik Müllers, Karen Akopyan, Kamila Burdova, Tobias Koolmeister, Thomas Helleday, René H Medema, Libor Macurek, Arne Lindqvist
After DNA damage, the cell cycle is arrested to avoid propagation of mutations. Arrest in G2 phase is initiated by ATM-/ATR-dependent signaling that inhibits mitosis-promoting kinases such as Plk1. At the same time, Plk1 can counteract ATR-dependent signaling and is required for eventual resumption of the cell cycle. However, what determines when Plk1 activity can resume remains unclear. Here, we use FRET-based reporters to show that a global spread of ATM activity on chromatin and phosphorylation of ATM targets including KAP1 control Plk1 re-activation...
June 12, 2017: EMBO Journal
https://www.readbyqxmd.com/read/28604853/a-single-quantum-dot-based-nanosensor-for-the-signal-on-detection-of-dna-methyltransferase
#18
Fei Ma, Wen-Jing Liu, Bo Tang, Chun-Yang Zhang
We develop a single quantum dot (QD)-based nanosensor for the signal-on detection of DNA methyltransferase (MTase). By integration of single-molecule counting with the QD-based fluorescence resonance energy transfer (FRET), the proposed nanosensor can sensitively detect DNA MTase with a detection limit of as low as 0.002 U mL(-1), and it can be further applied for inhibitor screening and accurate detection of DNA MTase in complex biological samples.
June 22, 2017: Chemical Communications: Chem Comm
https://www.readbyqxmd.com/read/28604851/analysis-of-ribonuclease-activity-in-sub-nanoliter-droplets-by-label-free-fluorescence-measurements
#19
Jae-Won Choi, Bala Murali Krishna Vasamsetti, Kyu-Wan Kim, Seung Hwan Seo, Dong-Hun Lee, Soo-Ik Chang, Jaebum Choo, Hak Yong Kim
We report the results of a label-free analysis of ribonuclease activity using droplet-based microfluidics. The ribonucleolytic activity of ribonucleases (RNases) plays a critical role in cellular functions such as development, survival, growth and differentiation. Altered ribonucleolytic activity and/or the expression level of the RNase A family are known to be associated with pancreatic, bladder, ovarian and thyroid cancers among others. For this reason, the RNase A family is a meaningful protein biomarker that can be used in the diagnosis of cancer and as a target for new drug screening...
June 12, 2017: Analyst
https://www.readbyqxmd.com/read/28604367/ultrafast-emission-decay-with-high-emission-efficiency-of-quantum-dots-in-plasmonic-dielectric-metasubstrates
#20
Waylin J Wing, Seyed M Sadeghi, Rithvik R Gutha
We report on the ultrafast decay of quantum dots that reaches nearly 430 ps without reduction in the emission intensity when near a metasubstrate of simple geometry. By implementing a layered structure of amorphous silicon sandwiched between two gold layers, the balance between plasmonic near field enhancement and energy transfer of the quantum dots has been shifted. This is achieved by tailoring the amount of Förster resonance energy transfer (FRET) to the top Au layer and plasmonic near field enhancement by the bottom Au layer...
June 12, 2017: Journal of Physics. Condensed Matter: An Institute of Physics Journal
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