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https://www.readbyqxmd.com/read/28431041/bidirectional-approaches-for-optogenetic-regulation-of-gene-expression-in-mammalian-cells-using-arabidopsis-cryptochrome-2
#1
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
#2
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
#3
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
#4
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...
April 19, 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
#5
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
#6
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
#7
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...
March 27, 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
#8
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
#9
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
#10
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...
March 8, 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
#11
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
#12
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
#13
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
https://www.readbyqxmd.com/read/28229215/sequence-analysis-and-expression-of-a-blue-light-photoreceptor-gene-slwc-1-from-the-cauliflower-mushroom-sparassis-latifolia
#14
Chi Yang, Lu Ma, Zhenghe Ying, Xiaoling Jiang, Yanquan Lin
Light is a necessary environmental factor for fruit body formation and development of the cauliflower mushroom Sparassis latifolia, a well-known edible and medicinal fungus. In this study, we firstly characterized the SP-C strain, which belonged to S. latifolia. And then we cloned and sequenced a photoreceptor gene (Slwc-1) from S. latifolia. The product of Slwc-1, SlWC-1 (872 aa residues) contained a coiled-coil region, a LOV domain, and two PAS domains. Phylogenetic tree result showed that SLWC-1 was most close to GfWC-1 from Grifola frondosa in edible and medicinal fungus...
April 2017: Current Microbiology
https://www.readbyqxmd.com/read/28211532/structure-of-a-lov-protein-in-apo-state-and-implications-for-construction-of-lov-based-optical-tools
#15
Vladimir Arinkin, Joachim Granzin, Katrin Röllen, Ulrich Krauss, Karl-Erich Jaeger, Dieter Willbold, Renu Batra-Safferling
Unique features of Light-Oxygen-Voltage (LOV) proteins like relatively small size (~12-19 kDa), inherent modularity, highly-tunable photocycle and oxygen-independent fluorescence have lately been exploited for the generation of optical tools. Structures of LOV domains reported so far contain a flavin chromophore per protein molecule. Here we report two new findings on the short LOV protein W619_1-LOV from Pseudomonas putida. First, the apo-state crystal structure of W619_1-LOV at 2.5 Å resolution reveals conformational rearrangements in the secondary structure elements lining the chromophore pocket including elongation of the Fα helix, shortening of the Eα-Fα loop and partial unfolding of the Eα helix...
February 17, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28194625/trypanosomiasis-challenge-estimation-using-the-diminazene-aceturate-berenil-index-in-zebu-in-gabon
#16
Brieuc G A Cossic, Brice Adjahoutonon, Pierre Gloaguen, Gui Lov Dibanganga, Gael Maganga, Pascal Leroy, Ewan T MacLeod, Kim Picozzi
A longitudinal study was conducted within a cattle ranch in Gabon to determine the diminazene aceturate (Berenil) index (DAI) in a group of Zebu, raised under low tsetse density; this measure providing an assessment of trypanosomiasis risk. The objective was to evaluate the trypanosomiasis pressure thus informing trypanosomiasis control methods and cattle management. Twenty female adult Zebu were monitored for 24 weeks during the dry season. Blood samples were collected on a weekly basis and subjected to parasitological and haematological analysis (n = 480), using the buffy-coat method and the packed cell volume value (PCV), respectively, infected animals were treated with a single intramuscular injection of diminazene aceturate (8 mg/kg)...
March 2017: Tropical Animal Health and Production
https://www.readbyqxmd.com/read/28145707/glutamine-amide-flip-elicits-long-distance-allosteric-responses-in-the-lov-protein-vivid
#17
Abir Ganguly, Walter Thiel, Brian R Crane
Light-oxygen-voltage (LOV) domains sense blue light through the photochemical formation of a cysteinyl-flavin covalent adduct. Concurrent protonation at the flavin N5 position alters the hydrogen bonding interactions of an invariant Gln residue that has been proposed to flip its amide side chain as a critical step in the propagation of conformational change. Traditional molecular dynamics (MD) and replica-exchange MD (REMD) simulations of the well-characterized LOV protein Vivid (VVD) demonstrate that the Gln182 amide indeed reorients by ∼180° in response to either adduct formation or reduction of the isoalloxazine ring to the neutral semiquinone, both of which involve N5 protonation...
February 13, 2017: Journal of the American Chemical Society
https://www.readbyqxmd.com/read/28127721/stabilization-and-amorphization-of-lovastatin-using-different-types-of-silica
#18
Mai Khanfar, Suhair Al-Nimry
Lovastatin (LOV), an antihyperlipidimic agent, is characterized by low solubility/poor dissolution and, thus, low bioavailability (<5%). A beneficial effect on its bioavailability could result from improving its dissolution. One of the most common methods used to enhance dissolution is the preparation of solid dispersions. Solid dispersions of LOV and silica with different surface areas were prepared. The effects of the type of silica, ratio of drug/silica, incubation period with silica, and the effect of surface area were all studied...
January 26, 2017: AAPS PharmSciTech
https://www.readbyqxmd.com/read/28100053/the-grateful-infrared-sequential-protein-structural-changes-resolved-by-infrared-difference-spectroscopy
#19
Tilman Kottke, Víctor A Lórenz-Fonfría, Joachim Heberle
The catalytic activity of proteins is a function of structural changes. Very often these are as minute as protonation changes, hydrogen bonding changes, and amino acid side chain reorientations. To resolve these, a methodology is afforded that not only provides the molecular sensitivity but allows for tracing the sequence of these hierarchical reactions at the same time. This feature article showcases results from time-resolved IR spectroscopy on channelrhodopsin (ChR), light-oxygen-voltage (LOV) domain protein, and cryptochrome (CRY)...
January 19, 2017: Journal of Physical Chemistry. B
https://www.readbyqxmd.com/read/28068090/femtosecond-to-millisecond-dynamics-of-light-induced-allostery-in-the-avena-sativa-lov-domain
#20
Agnieszka A Gil, Sergey P Laptenok, Jarrod B French, James N Iuliano, Andras Lukacs, Christopher R Hall, Igor V Sazanovich, Gregory M Greetham, Adelbert Bacher, Boris Illarionov, Markus Fischer, Peter J Tonge, Stephen R Meech
The rational engineering of photosensor proteins underpins the field of optogenetics, in which light is used for spatiotemporal control of cell signaling. Optogenetic elements function by converting electronic excitation of an embedded chromophore into structural changes on the microseconds to seconds time scale, which then modulate the activity of output domains responsible for biological signaling. Using time-resolved vibrational spectroscopy coupled with isotope labeling, we have mapped the structural evolution of the LOV2 domain of the flavin binding phototropin Avena sativa (AsLOV2) over 10 decades of time, reporting structural dynamics between 100 fs and 1 ms after optical excitation...
February 9, 2017: Journal of Physical Chemistry. B
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