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https://www.readbyqxmd.com/read/29113919/lighting-up-my-life-a-lov-based-fluorescent-reporter-for-campylobacter-jejuni
#1
Bassam A Elgamoudi, Julian M Ketley
In this study, a LOV-based fluorescent reporter (light, oxygen, or voltage-sensing domains of phototropin), termed iLOV, was adapted for Campylobacter jejuni and used to investigate promoter activity via monitoring fluorescence intensity and to study the localisation of two chemotaxis proteins. The pC46 complementation vector contains coding sequence from cj0046, a C. jejuni NCTC11168 pseudo-gene and is used to integrate cloned genes onto the C. jejuni chromosome. The pC46 vector was used to construct plasmids containing iLOV, driven by three different C...
November 4, 2017: Research in Microbiology
https://www.readbyqxmd.com/read/29042655/electron-transfer-pathways-in-a-light-oxygen-voltage-lov-protein-devoid-of-the-photoactive-cysteine
#2
Benita Kopka, Kathrin Magerl, Anton Savitsky, Mehdi D Davari, Katrin Röllen, Marco Bocola, Bernhard Dick, Ulrich Schwaneberg, Karl-Erich Jaeger, Ulrich Krauss
Blue-light absorption by the flavin chromophore in light, oxygen, voltage (LOV) photoreceptors triggers photochemical reactions that lead to the formation of a flavin-cysteine adduct. While it has long been assumed that adduct formation is essential for signaling, it was recently shown that LOV photoreceptor variants devoid of the photoactive cysteine can elicit a functional response and that flavin photoreduction to the neutral semiquinone radical is sufficient for signal transduction. Currently, the mechanistic basis of the underlying electron- (eT) and proton-transfer (pT) reactions is not well understood...
October 17, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28972909/solubility-and-transport-of-cr-iii-in-a-historically-contaminated-soil-evidence-of-a-rapidly-reacting-dimeric-cr-iii-organic-matter-complex
#3
Åsa Löv, Carin Sjöstedt, Mats Larsbo, Ingmar Persson, Jon Petter Gustafsson, Geert Cornelis, Dan B Kleja
Chromium is a common soil contaminant and, although it has been studied widely, questions about its speciation and dissolutions kinetics remain unanswered. We combined information from an irrigation experiment performed with intact soil columns with data from batch experiments to evaluate solubility and mobilization mechanisms of Cr(III) in a historically contaminated soil (>65 years). Particulate and colloidal Cr(III) forms dominated transport in this soil, but their concentrations were independent of irrigation intensity (2-20 mm h(-1))...
September 20, 2017: Chemosphere
https://www.readbyqxmd.com/read/28926259/dual-photochemical-reaction-pathway-in-flavin-based-photoreceptor-lov-domain-a-combined-quantum-mechanics-molecular-mechanics-investigation
#4
Setsuko Nakagawa, Oliver Weingart, Christel M Marian
The primary photochemical reaction of the light, oxygen, and voltage (LOV) domain of the blue-light photosensor YtvA of Bacillus subtilis were investigated using high-level QM(DFT/MRCI)/MM methods. After blue-light excitation, the Sγ atom of the reactive cysteine forms a covalent bond with the C4a of the flavin mononucleotide (FMN) ring. Two conformations for the side chain of reactive cysteine with occupancies of 70% (conf A) and 30% (conf B) are observed in the X-ray crystallographic structures of the YtvA-LOV ( Möglich , A...
October 4, 2017: Journal of Physical Chemistry. B
https://www.readbyqxmd.com/read/28893998/engineering-a-light-activated-caspase-3-for-precise-ablation-of-neurons-in-vivo
#5
Ashley D Smart, Roland A Pache, Nathan D Thomsen, Tanja Kortemme, Graeme W Davis, James A Wells
The circuitry of the brain is characterized by cell heterogeneity, sprawling cellular anatomy, and astonishingly complex patterns of connectivity. Determining how complex neural circuits control behavior is a major challenge that is often approached using surgical, chemical, or transgenic approaches to ablate neurons. However, all these approaches suffer from a lack of precise spatial and temporal control. This drawback would be overcome if cellular ablation could be controlled with light. Cells are naturally and cleanly ablated through apoptosis due to the terminal activation of caspases...
September 26, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28892628/no-photon-wasted-an-efficient-and-selective-singlet-oxygen-photosensitizing-protein
#6
Michael Westberg, Mikkel Bregnhøj, Michael Etzerodt, Peter R Ogilby
Optogenetics has been, and will continue to be, a boon to mechanistic studies of cellular processes. Genetically encodable proteins that sensitize the production of reactive oxygen species (ROS) are expected to play an increasingly important role, particularly in elucidating mechanisms of temporally and spatially dependent cell signaling. However, a substantial challenge in developing such photosensitizing proteins has been to funnel the optical excitation energy into the initial selective production of only one ROS...
September 27, 2017: Journal of Physical Chemistry. B
https://www.readbyqxmd.com/read/28889209/micro-bead-injection-spectroscopy-for-label-free-automated-determination-of-immunoglobulin-g-in-human-serum
#7
Inês I Ramos, Luís M Magalhães, Luisa Barreiros, Salette Reis, José L F C Lima, Marcela A Segundo
Immunoglobulin G (IgG) represents the major fraction of antibodies in healthy adult human serum, and deviations from physiological levels are a generic marker of disease corresponding to different pathologies. Therefore, screening methods for IgG evaluation are a valuable aid to diagnostics. The present work proposes a rapid, automatic, and miniaturized method based on UV-vis micro-bead injection spectroscopy (μ-BIS) for the real-time determination of human serum IgG with label-free detection. Relying on attachment of IgG in rec-protein G immobilized in Sepharose 4B, a bioaffinity column is automatically assembled, where IgG is selectively retained and determined by on-column optical density measurement...
September 9, 2017: Analytical and Bioanalytical Chemistry
https://www.readbyqxmd.com/read/28868541/photoinduced-dimerization-of-a-photosensory-dna-binding-protein-el222-and-its-lov-domain
#8
Akira Takakado, Yusuke Nakasone, Masahide Terazima
EL222 is a blue light sensor protein consisting of a light-oxygen-voltage (LOV) domain (EL-LOV domain) at the N-terminus and a helix-turn-helix DNA-binding domain at the C-terminus. EL222 acts as a light dependent transcriptional factor. The photochemical reactions of EL222 and the light sensing properties of the LOV domain were investigated. Concentration dependent experiments revealed that the EL-LOV domain is in equilibrium between the dimer and the monomer in the dark state, and the main photoreaction is the dimerization reaction between a monomer in the ground state and that in the excited state...
September 20, 2017: Physical Chemistry Chemical Physics: PCCP
https://www.readbyqxmd.com/read/28802576/atorvastatin-and-lovastatin-but-not-pravastatin-increased-cellular-complex-formation-between-pcsk9-and-the-ldl-receptor-in-human-hepatocyte-like-c3a-cells
#9
Quantil M Melendez, Catherine J Wooten, Dayami Lopez
Statins are the first-line treatment for hypercholesterolemic patients. Herein, the effects of three statins on complex formation between proprotein convertase subtilisin-kexin 9 (PCSK9) and the low density lipoprotein receptor (LDLR), a critical step for the PCSK9-dependent degradation of LDLR in the lysosome, were examined. Human hepatocyte-like C3A cells grown in control (containing 10% fetal bovine serum) or MITO+ (supplemented with BD™ MITO + serum extender) medium were also treated with atorvastatin (Atorv), lovastatin (Lov), or pravastatin (Prav) for 24 h...
October 7, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/28797596/an-update-on-aureochromes-phylogeny-mechanism-function
#10
REVIEW
Peter G Kroth, Christian Wilhelm, Tilman Kottke
Light is important for algae, as it warrants metabolic independence via photosynthesis. In addition to the absorption of light by the photosystems, algae possess a variety of specific photoreceptors that allow the quantification of the light fluxes as well as the assessment of light qualities. About a decade ago, aureochromes have been described in the xanthophyte alga Vaucheria frigida. These proteins represent a new type of blue light photoreceptor as they possess both a light-oxygen-voltage (LOV) domain for light reception as well as a basic region leucine zipper (bZIP) domain for DNA binding, indicating that they represent light-driven transcription factors...
July 11, 2017: Journal of Plant Physiology
https://www.readbyqxmd.com/read/28680190/a-labview-%C3%A2-based-software-for-the-control-of-the-autorad-platform-a-fully-automated-multisequential-flow-injection-analysis-lab-on-valve-msfia-lov-system-for-radiochemical-analysis
#11
Donato Barbesi, Víctor Vicente Vilas, Sylvain Millet, Miguel Sandow, Jean-Yves Colle, Laura Aldave de Las Heras
A LabVIEW(®)-based software for the control of the fully automated multi-sequential flow injection analysis Lab-on-Valve (MSFIA-LOV) platform AutoRAD performing radiochemical analysis is described. The analytical platform interfaces an Arduino(®)-based device triggering multiple detectors providing a flexible and fit for purpose choice of detection systems. The different analytical devices are interfaced to the PC running LabVIEW(®)VI software using USB and RS232 interfaces, both for sending commands and receiving confirmation or error responses...
2017: Journal of Radioanalytical and Nuclear Chemistry
https://www.readbyqxmd.com/read/28663371/functional-characterization-of-a-constitutively-active-kinase-variant-of-arabidopsis-phototropin-1
#12
COMPARATIVE STUDY
Jan Petersen, Shin-Ichiro Inoue, Sharon M Kelly, Stuart Sullivan, Toshinori Kinoshita, John M Christie
Phototropins (phots) are plasma membrane-associated serine/threonine kinases that coordinate a range of processes linked to optimizing photosynthetic efficiency in plants. These photoreceptors contain two light-, oxygen-, or voltage-sensing (LOV) domains within their N terminus, with each binding one molecule of flavin mononucleotide as a UV/blue light-absorbing chromophore. Although phots contain two LOV domains, light-induced activation of the C-terminal kinase domain and subsequent receptor autophosphorylation is controlled primarily by the A'α-LOV2-Jα photosensory module...
August 18, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28632317/quantum-mechanics-molecular-mechanics-study-on-the-photoreactions-of-dark-and-light-adapted-states-of-a-blue-light-ytva-lov-photoreceptor
#13
Xue-Ping Chang, Yuan-Jun Gao, Wei-Hai Fang, Ganglong Cui, Walter Thiel
The dark- and light-adapted states of YtvA LOV domains exhibit distinct excited-state behavior. We have employed high-level QM(MS-CASPT2)/MM calculations to study the photochemical reactions of the dark- and light-adapted states. The photoreaction from the dark-adapted state starts with an S1 →T1 intersystem crossing followed by a triplet-state hydrogen transfer from the thiol to the flavin moiety that produces a diradical intermediate, and a subsequent internal conversion that triggers a barrierless C-S bond formation in the S0 state...
August 1, 2017: Angewandte Chemie
https://www.readbyqxmd.com/read/28576317/a-miniature-optical-emission-spectrometric-system-in-a-lab-on-valve-for-sensitive-determination-of-cadmium
#14
Ya-Jie Zhang, Yi Cai, Yong-Liang Yu, Jian-Hua Wang
A miniature optical emission spectrometric (OES) system is developed by incorporating dielectric barrier discharge (DBD) microplasma as excitation source in a lab-on-valve (LOV) configuration for trace cadmium analysis. The entire system integrates bead injection (BI) sample pretreatment, post-column derivatization, in-situ nebulization and OES detection. Trace cadmium in sample solution is separated and preconcentrated by flowing through a 2-(5-Br-2-pyridylazo)-5-dethylaminophenol (5-Br-PADAP) loaded microcolumn in LOV, followed by elution with 20 μL of 0...
July 11, 2017: Analytica Chimica Acta
https://www.readbyqxmd.com/read/28543056/cloning-and-analysis-of-the-glwc-1-and-glwc-2-genes-encoding-putative-blue-light-photoreceptor-from-ganoderma-lucidum
#15
Xinran Xu, Xiangdong Chen, Wumengxiao Yu, Yu Liu, Weiwei Zhang, Jin Lan
Blue light plays an important role during the growth of Ganoderma lucidum, one of the best-known medicinal macrofungi in China. In the present study, we cloned Glwc-1 and Glwc-2, the homologue of the blue light photoreceptors Ncwc-1 and Ncwc-2 of Neurospora crassa, from G. lucidum. The deduced amino acid sequence of Glwc-1 contained the similar function domains as NcWC-1 including LOV, PAS B, PAS C, and PAC domains. The deduced amino acid sequence of Glwc-2 contained PAS domain and GATA-type zinc finger (Znf) domain as well as NcWC-2...
August 2017: Journal of Basic Microbiology
https://www.readbyqxmd.com/read/28530801/hydrogen-bonding-environment-of-the-n3-h-group-of-flavin-mononucleotide-in-the-light-oxygen-voltage-domains-of-phototropins
#16
Tatsuya Iwata, Dai Nozaki, Atsushi Yamamoto, Takayuki Koyama, Yasuzo Nishina, Kiyoshi Shiga, Satoru Tokutomi, Masashi Unno, Hideki Kandori
The light oxygen voltage (LOV) domain is a flavin-binding blue-light receptor domain, originally found in a plant photoreceptor phototropin (phot). Recently, LOV domains have been used in optogenetics as the photosensory domain of fusion proteins. Therefore, it is important to understand how LOV domains exhibit light-induced structural changes for the kinase domain regulation, which enables the design of LOV-containing optogenetics tools with higher photoactivation efficiency. In this study, the hydrogen bonding environment of the N3-H group of flavin mononucleotide (FMN) of the LOV2 domain from Adiantum neochrome (neo) 1 was investigated by low-temperature Fourier transform infrared spectroscopy...
June 20, 2017: Biochemistry
https://www.readbyqxmd.com/read/28502782/time-resolved-x-ray-solution-scattering-reveals-the-structural-photoactivation-of-a-light-oxygen-voltage-photoreceptor
#17
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...
June 6, 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
#18
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
#19
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
#20
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
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