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https://www.readbyqxmd.com/read/28530801/hydrogen-bonding-environment-of-the-n3-h-group-of-fmn-in-the-lov-domains-of-phototropins
#1
Tatsuya Iwata, Dai Nozaki, Atsushi Yamamoto, Takayuki Koyama, Yasuzo Nishina, Kiyoshi Shiga, Satoru Tokutomi, Masashi Unno, Hideki Kandori
LOV (light-oxygen-voltage) domain is a flavin-binding blue-light receptor domain originally found in plant phototropin, a photoreceptor for phototropism, chloroplast movement and stomata opening. LOV domains are available as optogenetics tools for photosensory domain of fusion proteins. Therefore, it is important to investigate how LOV domains show light-induced structural changes not only for the elucidation of regulation mechanism of kinase domain but also for the appropriate activation of other functional domains by LOV domain...
May 22, 2017: Biochemistry
https://www.readbyqxmd.com/read/28502782/time-resolved-x-ray-solution-scattering-reveals-the-structural-photoactivation-of-a-light-oxygen-voltage-photoreceptor
#2
Oskar Berntsson, Ralph P Diensthuber, Matthijs R Panman, Alexander Björling, Ashley J Hughes, Léocadie Henry, Stephan Niebling, Gemma Newby, Marianne Liebi, Andreas Menzel, Robert Henning, Irina Kosheleva, Andreas Möglich, Sebastian Westenhoff
Light-oxygen-voltage (LOV) receptors are sensory proteins controlling a wide range of organismal adaptations in multiple kingdoms of life. Because of their modular nature, LOV domains are also attractive for use as optogenetic actuators. A flavin chromophore absorbs blue light, forms a bond with a proximal cysteine residue, and induces changes in the surroundings. There is a gap of knowledge on how this initial signal is relayed further through the sensor to the effector module. To characterize these conformational changes, we apply time-resolved X-ray scattering to the homodimeric LOV domain from Bacillus subtilis YtvA...
May 8, 2017: Structure
https://www.readbyqxmd.com/read/28501154/new-approach-to-h-point-standard-addition-method-for-detection-and-elimination-of-unspecific-interferences-in-samples-with-unknown-matrix
#3
Marcin Wieczorek, Silvia Rengevicova, Paweł Świt, Aneta Woźniakiewicz, Joanna Kozak, Paweł Kościelniak
A novel method for correction of unknown unspecific (additive) interferences was developed in sequential injection analysis (SIA) using a Lab on Valve module (LOV) with spectrophotometric detection. The method implements a novel idea to calibrate by the standard addition method in several different chemical conditions created in such a way to measure different signal for an analyte and unchanged signal for interferents causing additive effect. This approach, being an enhancement of the H-point standard addition method (HPSAM), enables to quantify unbiased concentration of an analyte in the presence of unknown interferences...
August 1, 2017: Talanta
https://www.readbyqxmd.com/read/28500719/novel-thermostable-flavin-binding-fluorescent-proteins-from-thermophilic-organisms
#4
Marcus Wingen, Karl-Erich Jaeger, Thomas Gensch, Thomas Drepper
Flavin-binding fluorescent proteins (FbFPs) are small, oxygen-independent in vivo reporters, derived from Light Oxygen Voltage (LOV) domains of photoreceptors. Here, we investigated the thermostability of existing, as well as novel FbFPs, whose genes were identified in genome sequences of various thermophilic bacteria as well as metagenomic libraries from hot springs in the Yellowstone National Park. Detailed in vitro analyses revealed that two of those fluorescent reporter proteins were highly thermostable, exhibiting melting temperatures above 75°C...
May 2017: Photochemistry and Photobiology
https://www.readbyqxmd.com/read/28500710/characterization-of-the-blue-light-activated-adenylyl-cyclase-mpac-by-flash-photolysis-and-ftir-spectroscopy
#5
Silke Kerruth, Pit Langner, Sarah Raffelberg, Wolfgang Gärtner, Joachim Heberle
The recently discovered photo-activated adenylyl cyclase (mPAC from Microcoleus chthonoplastes) is the first PAC that owes a light-, oxygen- and voltage-sensitive (LOV) domain for blue-light sensing. The photoreaction of the mPAC receptor was studied by time-resolved UV/vis and light-induced Fourier transform infrared (FTIR) absorption difference spectroscopy. The photocycle comprises of the typical triplet state LOV715 and the thio-adduct state LOV390 . While the adduct state decays with a time constant of 8 s, the lifetime of the triplet state is with 656 ns significantly shorter than in all other reported LOV domains...
May 2017: Photochemistry and Photobiology
https://www.readbyqxmd.com/read/28478682/fully-automated-system-for-99-tc-monitoring-in-hospital-and-urban-residues-a-simple-approach-to-waste-management
#6
Marina Villar, Antoni Borràs, Jessica Avivar, Fernando Vega, Víctor Cerdà, Laura Ferrer
(99)Tc is an artificial beta emitter widely used in nuclear medicine for diagnostic tests. A fully automated and rapid system for (99)Tc monitoring is introduced with the aim to improve hospital residues management. This system can also be helpful for controlling urban wastewater. Figures of merit similar to those obtained with liquid scintillation counting were achieved by exploiting a simple, economic, and portable system with spectrophotometric detection. The combination of flow analysis techniques, i.e...
May 16, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28469162/blue-light-reception-through-quaternary-transitions
#7
Christopher Engelhard, Ralph P Diensthuber, Andreas Möglich, Robert Bittl
Sensory photoreceptors absorb light via their photosensor modules and trigger downstream physiological adaptations via their effector modules. Light reception accordingly depends on precisely orchestrated interactions between these modules, the molecular details of which often remain elusive. Using electron-electron double resonance (ELDOR) spectroscopy and site-directed spin labelling, we chart the structural transitions facilitating blue-light reception in the engineered light-oxygen-voltage (LOV) histidine kinase YF1 which represents a paradigm for numerous natural signal receptors...
May 3, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28431041/bidirectional-approaches-for-optogenetic-regulation-of-gene-expression-in-mammalian-cells-using-arabidopsis-cryptochrome-2
#8
Gopal P Pathak, Jessica I Spiltoir, Camilla Höglund, Lauren R Polstein, Sari Heine-Koskinen, Charles A Gersbach, Jari Rossi, Chandra L Tucker
Optogenetic tools allow regulation of cellular processes with light, which can be delivered with spatiotemporal resolution. In previous work, we used cryptochrome 2 (CRY2) and CIB1, Arabidopsis proteins that interact upon light illumination, to regulate transcription with light in yeast. While adopting this approach to regulate transcription in mammalian cells, we observed light-dependent redistribution and clearing of CRY2-tethered proteins within the nucleus. The nuclear clearing phenotype was dependent on the presence of a dimerization domain contained within the CRY2-fused transcriptional activators...
April 20, 2017: Nucleic Acids Research
https://www.readbyqxmd.com/read/28425502/revealing-hidden-conformational-space-of-lov-protein-vivid-through-rigid-residue-scan-simulations
#9
Hongyu Zhou, Brian D Zoltowski, Peng Tao
VIVID(VVD) protein is a Light-Oxygen-Voltage(LOV) domain in circadian clock system. Upon blue light activation, a covalent bond is formed between VVD residue Cys108 and its cofactor flavin adenine dinucleotide(FAD), and prompts VVD switching from Dark state to Light state with significant conformational deviation. However, the mechanism of this local environment initiated global protein conformational change remains elusive. We employed a recently developed computational approach, rigid residue scan(RRS), to systematically probe the impact of the internal degrees of freedom in each amino acid residue of VVD on its overall dynamics by applying rigid body constraint on each residue in molecular dynamics simulations...
April 20, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28412349/light-induced-protein-degradation-in-human-derived-cells
#10
Wansheng Sun, Wenyao Zhang, Chao Zhang, Miaowei Mao, Yuzheng Zhao, Xianjun Chen, Yi Yang
Controlling protein degradation can be a valuable tool for posttranslational regulation of protein abundance to study complex biological systems. In the present study, we designed a light-switchable degron consisting of a light oxygen voltage (LOV) domain of Avena sativa phototropin 1 (AsLOV2) and a C-terminal degron. Our results showed that the light-switchable degron could be used for rapid and specific induction of protein degradation in HEK293 cells by light in a proteasome-dependent manner. Further studies showed that the light-switchable degron could also be utilized to mediate the degradation of secreted Gaussia princeps luciferase (GLuc), demonstrating the adaptability of the light-switchable degron in different types of protein...
April 12, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/28387114/light-induced-conformational-changes-of-lov2-kinase-and-the-linker-region-in-arabidopsis-phototropin2
#11
Akira Takakado, Yusuke Nakasone, Koji Okajima, Satoru Tokutomi, Masahide Terazima
Phototropins (phots) are blue light sensors found in a variety of higher plants and algae. The photochemical reactions of this family of proteins have attracted much attention since their discovery. Phots have two light sensor domains called light-oxygen-voltage 1 (LOV1) and LOV2. After the formation of the characteristic adduct of the LOV domain, a conformational change of the C-terminal region of the LOV2 domain occurs, and characterizing this change is important for understanding biological function, that is, kinase activation...
May 4, 2017: Journal of Physical Chemistry. B
https://www.readbyqxmd.com/read/28378460/chlorosis-caused-by-two-recessively-interacting-genes-reveals-a-role-of-rna-helicase-in-hybrid-breakdown-in-arabidopsis-thaliana
#12
Björn Plötner, Markus Nurmi, Axel Fischer, Mutsumi Watanabe, Korbinian Schneeberger, Svante Holm, Neha Vaid, Mark Aurel Schöttler, Dirk Walther, Rainer Hoefgen, Detlef Weigel, Roosa A E Laitinen
Hybrids often differ in fitness from their parents. They may be superior, translating into hybrid vigour or heterosis, but they may also be markedly inferior, because of hybrid weakness or incompatibility. The underlying genetic causes for the latter can often be traced back to genes that evolve rapidly because of sexual or host-pathogen conflicts. Hybrid weakness may manifest itself only in later generations, in a phenomenon called hybrid breakdown. We have characterized a case of hybrid breakdown among two Arabidopsis thaliana accessions, Shahdara (Sha, Tajikistan) and Lövvik-5 (Lov-5, Northern Sweden)...
April 4, 2017: Plant Journal: for Cell and Molecular Biology
https://www.readbyqxmd.com/read/28369414/the-european-lead-extraction-controlled-electra-study-a-european-heart-rhythm-association-ehra-registry-of-transvenous-lead-extraction-outcomes
#13
Maria Grazia Bongiorni, Charles Kennergren, Christian Butter, Jean Claude Deharo, Andrzej Kutarski, Christopher A Rinaldi, Simone L Romano, Aldo P Maggioni, Maryna Andarala, Angelo Auricchio, Karl-Heinz Kuck, Carina Blomström-Lundqvist
Aims: The European Lead Extraction ConTRolled Registry (ELECTRa), is a prospective registry of consecutive transvenous lead extraction (TLE) procedures conducted by the European Heart Rhythm Association (EHRA) in order to identify the safety and efficacy of the current practice of TLE. Methods and results: European centres performing TLE, invited by the organizing committee on behalf of EHRA, prospectively recruited all consecutive patients undergoing TLE at their institution...
March 23, 2017: European Heart Journal
https://www.readbyqxmd.com/read/28346403/near-infrared-optogenetic-pair-for-protein-regulation-and-spectral-multiplexing
#14
Taras A Redchuk, Evgeniya S Omelina, Konstantin G Chernov, Vladislav V Verkhusha
Multifunctional optogenetic systems are in high demand for use in basic and biomedical research. Near-infrared-light-inducible binding of bacterial phytochrome BphP1 to its natural PpsR2 partner is beneficial for simultaneous use with blue-light-activatable tools. However, applications of the BphP1-PpsR2 pair are limited by the large size, multidomain structure and oligomeric behavior of PpsR2. Here, we engineered a single-domain BphP1 binding partner, Q-PAS1, which is three-fold smaller and lacks oligomerization...
June 2017: Nature Chemical Biology
https://www.readbyqxmd.com/read/28340699/a-led-based-fiber-optic-sensor-integrated-with-lab-on-valve-manifold-for-colorimetric-determination-of-free-chlorine-in-water
#15
Yan Xiong, Jun Tan, Shenwen Fang, Chengjie Wang, Qing Wang, Jiayi Wu, Jie Chen, Ming Duan
In this work, a colorimetric methodology was developed for free chlorine determination by constructing a fiber-optic sensor (FOS) which was designed by using a small LED as light source and two silica fibers as light transmission elements. Then the FOS was integrated with the sequential injection lab-on-valve (SI-LOV) manifold to fabricate a FOS-SI-LOV system for free chlorine detection. The colorimetric determination was based on the reaction between N,N-diethyl-p-phenylenediamine (DPD) reagent and free chlorine, which would produce a chromophore showing maximum absorbance at 511 and 551nm and detected by a micro spectrometer detector...
May 15, 2017: Talanta
https://www.readbyqxmd.com/read/28293891/development-of-a-synthetic-switch-to-control-protein-stability-in-eukaryotic-cells-with-light
#16
Christof Taxis
In eukaryotic cells, virtually all regulatory processes are influenced by proteolysis. Thus, synthetic control of protein stability is a powerful approach to influence cellular behavior. To achieve this, selected target proteins are modified with a conditional degradation sequence (degron) that responds to a distinct signal. For development of a synthetic degron, an appropriate sensor domain is fused with a degron such that activity of the degron is under control of the sensor. This chapter describes the development of a light-activated, synthetic degron in the model organism Saccharomyces cerevisiae...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28271102/switching-from-adduct-formation-to-electron-transfer-in-a-light-oxygen-voltage-domain-containing-the-reactive-cysteine
#17
Kathrin Magerl, Ivan Stambolic, Bernhard Dick
LOV (light-, oxygen- or voltage-sensitive) domains act as photosensory units of many prokaryotic and eukaryotic proteins. Upon blue light excitation they undergo a photocycle via the excited triplet state of their flavin chromophore yielding the flavin-cysteinyl adduct. Adduct formation is highly conserved among all LOV domains and constitutes the primary step of LOV domain signaling. But recently, it has been shown that signal propagation can also be triggered by flavin photoreduction to the neutral semiquinone offering new prospects for protein engineering...
May 3, 2017: Physical Chemistry Chemical Physics: PCCP
https://www.readbyqxmd.com/read/28259481/lung-protective-ventilation-initiated-in-the-emergency-department-lov-ed-a%C3%A2-quasi-experimental-before-after-trial
#18
Brian M Fuller, Ian T Ferguson, Nicholas M Mohr, Anne M Drewry, Christopher Palmer, Brian T Wessman, Enyo Ablordeppey, Jacob Keeperman, Robert J Stephens, Cristopher C Briscoe, Angelina A Kolomiets, Richard S Hotchkiss, Marin H Kollef
STUDY OBJECTIVE: We evaluated the efficacy of an emergency department (ED)-based lung-protective mechanical ventilation protocol for the prevention of pulmonary complications. METHODS: This was a quasi-experimental, before-after study that consisted of a preintervention period, a run-in period of approximately 6 months, and a prospective intervention period. The intervention was a multifaceted ED-based mechanical ventilator protocol targeting lung-protective tidal volume, appropriate setting of positive end-expiratory pressure, rapid oxygen weaning, and head-of-bed elevation...
March 1, 2017: Annals of Emergency Medicine
https://www.readbyqxmd.com/read/28257211/temperature-sensitive-singlet-oxygen-photosensitization-by-lov-derived-fluorescent-flavoproteins
#19
Michael Westberg, Mikkel Bregnhøj, Michael Etzerodt, Peter R Ogilby
Optogenetic sensitizers that selectively produce a given reactive oxygen species (ROS) constitute a promising tool for studying cell signaling processes with high levels of spatiotemporal control. However, to harness the full potential of this tool for live cell studies, the photophysics of currently available systems need to be explored further and optimized. Of particular interest in this regard, are the flavoproteins miniSOG and SOPP, both of which (1) contain the chromophore flavin mononucleotide, FMN, in a LOV-derived protein enclosure, and (2) photosensitize the production of singlet oxygen, O2(a(1)Δg)...
March 16, 2017: Journal of Physical Chemistry. B
https://www.readbyqxmd.com/read/28244872/kinetics-of-the-lov-domain-of-zeitlupe-determine-its-circadian-function-in-arabidopsis
#20
Ashutosh Pudasaini, Jae Sung Shim, Young Hun Song, Hua Shi, Takatoshi Kiba, David E Somers, Takato Imaizumi, Brian D Zoltowski
A LOV (Light, Oxygen, or Voltage) domain containing blue-light photoreceptor ZEITLUPE (ZTL) directs circadian timing by degrading clock proteins in plants. Functions hinge upon allosteric differences coupled to the ZTL photocycle; however, structural and kinetic information was unavailable. Herein, we tune the ZTL photocycle over two orders of magnitude. These variants reveal that ZTL complexes with targets independent of light, but dictates enhanced protein degradation in the dark. In vivo experiments definitively show photocycle kinetics dictate the rate of clock component degradation, thereby impacting circadian period...
February 28, 2017: ELife
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