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Fluorescent protein

Nawel Haïli, Julien Louap, Michel Canonge, Franjo Jagic, Christelle Louis-Mondésir, Thierry Chardot, Pierre Briozzo
The membrane proteins acyl-CoA:diacylglycerol acyltransferases (DGAT) are essential actors for triglycerides (TG) biosynthesis in eukaryotic organisms. Microbial production of TG is of interest for producing biofuel and value-added novel oils. In the oleaginous yeast Yarrowia lipolytica, Dga1p enzyme from the DGAT2 family plays a major role in TG biosynthesis. Producing recombinant DGAT enzymes pure and catalytically active is difficult, hampering their detailed functional characterization. In this report, we expressed in Escherichia coli and purified two soluble and active forms of Y...
2016: PloS One
Zaiddodine Pashandi, Maryam Molakarimi, Reza H Sajedi, Majid Taghdir, Hossein Naderi-Manesh
Mnemiopsin, an EF-hand Ca(2+) binding photoprotein isolated from luminous ctenophore Mnemiopsis leidyi, emits blue light from its chromophore, coelenterazine, which is non-covalently bond in its central hydrophobic core. Previous studies have revealed unique biochemical properties for ctenophore photoproteins such as inactivation by light, but only few have focused on photoinactivation process. To understand the nature of photoinactivation process we have investigated the impact of light alone and in the presence of Ca(2+) ion on the structure of this photoprotein...
October 18, 2016: Journal of Photochemistry and Photobiology. B, Biology
Robert T Dauchy, Melissa A Wren-Dail, Aaron E Hoffman, John P Hanifin, Benjamin Warfield, George C Brainard, Steven M Hill, Victoria P Belancio, Erin M Dauchy, David E Blask
Regular cycles of exposure to light and dark control pineal melatonin production and temporally coordinate circadian rhythms of metabolism and physiology in mammals. Previously we demonstrated that the peak circadian amplitude of nocturnal blood melatonin levels of rats were more than 6-fold higher after exposure to cool white fluorescent (CWF) light through blue-tinted (compared with clear) rodent cages. Here, we evaluated the effects of light-phase exposure of rats to white light-emitting diodes (LED), which emit light rich in the blue-appearing portion of the visible spectrum (465-485 nm), compared with standard broadspectrum CWF light, on melatonin levels during the subsequent dark phase and on plasma measures of metabolism and physiology...
2016: Comparative Medicine
Enrico Condemi, Denis Prim, Rebecca Brönnimann, Jonathan Fusco, Simon Crelier, Olimpia Mamula Steiner, Jean-Manuel Segura
No abstract text is available yet for this article.
October 2016: Chimia
Oliver R Oakley, Kee Jun Kim, Po-Ching Lin, Radwa Barakat, Joseph A Cacioppo, Zhong Li, Alex Whitaker, Kwang Chul Chung, Wenyan Mei, CheMyong Ko
17β-estradiol is a potent sex hormone synthesized primarily by gonads in females and males that regulates development and function of the reproductive system. Recent studies show that 17β-estradiol is locally synthesized in non-reproductive tissues and regulates a myriad of events, including local inflammatory responses. In this study, we report that mesenteric lymph nodes and Peyer's patches are novel sites of de novo synthesis of 17β-estradiol. These secondary lymphoid organs are located within or close to the gastrointestinal tract, contain leukocytes, and function at the forefront of immune surveillance...
October 25, 2016: Endocrinology
Zheng Zhang, Hannah A Ledford, Seojin Park, Wenying Wang, Sassan Rafizadeh, Hyo Jeong Kim, Wilson Xu, Ling Lu, Victor C Lau, Anne A Knowlton, Xiao-Dong Zhang, Ebenezer N Yamoah, Nipavan Chiamvimonvat
The normal function of ion channels depends critically on the precise subcellular localization and the number of channel proteins on the cell surface membrane. Small-conductance, Ca(2+) -activated K(+) channels (SK, KCa 2) are expressed in human atrial myocytes and responsible for shaping atrial action potentials. Understanding the mechanisms of SK channel trafficking may provide new insights into the regulation controlling the repolarization of atrial myocytes. We have previously demonstrated that the C and N termini of SK2 channels interact with actin-binding proteins, α-actinin2 and filamin A, respectively...
October 25, 2016: Journal of Physiology
Li Zhang, Wei Song, Ru-Ping Liang, Jian-Ding Qiu
We design two artificial substrate peptides to synthesize blue-emissive Cu nanoclusters and red-emissive Au nanoclusters, respectively. In addition to the biomineralization function, these two peptides retain the biological activity to be phosphorylated by protein kinase and digested by carboxypeptidase Y. In the absence of protein kinase, the peptides capped on the nanoclusters suffer consecutive exocleavage by carboxypeptidase Y, resulting in oxidation and thus fluorescence quenching of the nanoclusters due to the losing of peptide protection...
October 25, 2016: Analytical Chemistry
Shalev Gihaz, Diána Weiser, Adi Dror, Péter Sátorhelyi, Moran Jerabek-Willemsen, László Poppe, Ayelet Fishman
Two ternary sol-gel matrices, an octyltriethoxysilane-based aliphatic matrix and a phenyltriethoxysilane (PTEOS)-based aromatic matrix, were used to immobilize a methanol-stable variant of lipase from Geobacillus stearothermophilus T6 for the synthesis of biodiesel from waste oil. Superior thermal stability of the mutant versus the wildtype in methanol was confirmed by intrinsic protein fluorescence measurements. The influence of skim milk and soluble E. coli lysate proteins as bulking and stabilizing agents in conjunction with sol-gel entrapment were investigated...
October 25, 2016: ChemSusChem
Guillem Paniagua Soriano, Herman S Overkleeft, Bogdan I Florea
Activity-based protein profiling (ABPP) is a method to highlight enzymatic activities in a biological sample, which uses chemical probes that react covalently with the catalytic nucleophile of the enzyme. To circumvent disadvantages associated with the presence of reporter tags on chemical probes, the probe is equipped with a ligation handle to which the reporter can be reacted at the desired time and place in the ABPP workflow. This chapter demonstrates the power of a triple bioorthogonal ligation strategy which addresses the three activities of the proteasome: the β5-subunit selective norbornene-tagged probe is reacted with fluorescent tetrazine, the β1-selective azide-functionalized probe was addressed with a biotinylated phosphine, followed by an alkyne-substituted pan-reactive probe to label the remaining β2 activity to which an azide-coupled fluorophore was ligated...
2017: Methods in Molecular Biology
Marc P Baggelaar, Mario Van der Stelt
Competitive activity-based protein profiling is a highly efficient chemical biology technique to determine target engagement and selectivity profiles of enzyme inhibitors in complex proteomes. Fluorophosphonate-based fluorescent inhibitors are widely used as broad-spectrum probes for serine hydrolases. However, diacylglycerol lipase-α is not labeled by fluorophosphonate-based probes. To overcome this problem, we have developed a tailor-made activity-based probe that reacts with diacylglycerol lipase-α. Here we describe a case study in which we apply competitive activity-based protein profiling using a broad-spectrum and a tailor-made activity-based probe to establish selectivity and activity profiles of inhibitors targeting diacylglycerol lipase-α in the mouse brain proteome...
2017: Methods in Molecular Biology
Chien-Yuan Lin, Joseph E Jakes, Bryon S Donohoe, Peter N Ciesielski, Haibing Yang, Sophie-Charlotte Gleber, Stefan Vogt, Shi-You Ding, Wendy A Peer, Angus S Murphy, Maureen C McCann, Michael E Himmel, Melvin P Tucker, Hui Wei
BACKGROUND: Plant lignocellulosic biomass is an abundant, renewable feedstock for the production of biobased fuels and chemicals. Previously, we showed that iron can act as a co-catalyst to improve the deconstruction of lignocellulosic biomass. However, directly adding iron catalysts into biomass prior to pretreatment is diffusion limited, and increases the cost of biorefinery operations. Recently, we developed a new strategy for expressing iron-storage protein ferritin intracellularly to accumulate iron as a catalyst for the downstream deconstruction of lignocellulosic biomass...
2016: Biotechnology for Biofuels
Sunny C Y Jeng, Hedy H Y Chan, Evan P Booy, Sean A McKenna, Peter J Unrau
The effective tracking and purification of biological RNAs and RNA protein complexes is currently challenging. One promising strategy to simultaneously address both of these problems is to develop high-affinity RNA aptamers against taggable small molecule fluorophores. RNA Mango is a 39-nucleotide, parallel-stranded G-quadruplex RNA aptamer motif that binds with nanomolar affinity to a set of thiazole orange (TO1) derivatives while simultaneously inducing a 10(3)-fold increase in fluorescence. We find that RNA Mango has a large increase in its thermal stability upon the addition of its TO1-Biotin ligand...
October 24, 2016: RNA
Graziano Colombo, Marco Clerici, Alessandra Altomare, Francesco Rusconi, Daniela Giustarini, Nicola Portinaro, Maria Lisa Garavaglia, Ranieri Rossi, Isabella Dalle-Donne, Aldo Milzani
: In this study, we assessed the oxidative damage occurring in plasma proteins when human blood was exposed to inflammatory concentrations of hypochlorous acid (HOCl). We used specific thiol labelling and Western blot analyses to determine protein thiol oxidation, as well as analytical gel filtration HPLC coupled to fluorescence detection to explore formation of high molecular weight (HMW) protein aggregates. Thiol-containing proteins oxidized by HOCl were identified by redox proteomics...
October 21, 2016: Journal of Proteomics
Fei Ma, Xin Zhang, Xi Zhu, Tianpei Li, Jiao Zhan, Hui Chen, Chenliu He, Qiang Wang
Iron stress-induced protein A (IsiA) is a major chlorophyll-binding protein in the thylakoid membrane that is significantly induced under iron deficiency conditions. Using immunoblot analysis and 77k fluorescence spectroscopy combined with sucrose gradient fractionation, we monitored dynamic changes of IsiA-containing complexes in Synechocystis sp. PCC 6803 during exposure to long-term iron deficiency. Within 3 days of exposure to iron deficiency conditions, the initially induced free IsiA proteins preferentially conjugated to PS I trimer to form IsiA18-PS I trimers, which serve as light energy collectors for efficiently transmitting energy to PS I...
October 21, 2016: Molecular Plant
Daniel P Brink, Celina Borgström, Felipe G Tueros, Marie F Gorwa-Grauslund
BACKGROUND: The sugar sensing and carbon catabolite repression in Baker's yeast Saccharomyces cerevisiae is governed by three major signaling pathways that connect carbon source recognition with transcriptional regulation. Here we present a screening method based on a non-invasive in vivo reporter system for real-time, single-cell screening of the sugar signaling state in S. cerevisiae in response to changing carbon conditions, with a main focus on the response to glucose and xylose. RESULTS: The artificial reporter system was constructed by coupling a green fluorescent protein gene (yEGFP3) downstream of endogenous yeast promoters from the Snf3p/Rgt2p, SNF1/Mig1p and cAMP/PKA signaling pathways: HXT1p/2p/4p; SUC2p, CAT8p; TPS1p/2p and TEF4p respectively...
October 24, 2016: Microbial Cell Factories
Wipada Duangprompo, Kalaya Aree, Arunporn Itharat, Pintusorn Hansakul
5,6-dihydroxy-2,4-dimethoxy-9,10-dihydrophenanthrene (HMP) is an active compound isolated from the rhizome extracts of Dioscorea membranacea Pierre, a Thai medicinal plant. This study aimed to investigate the growth-inhibitory and apoptosis-inducing effects of HMP in human lung cancer A549 cells. The antiproliferative and cytotoxic effects of HMP were analyzed by a Sulforhodamine B assay. Cell division, cell cycle distribution and membrane asymmetry changes were each performed with different fluorescent dyes and then analyzed by flow cytometry...
October 25, 2016: American Journal of Chinese Medicine
Iuliana Aprodu, Florentina-Mihaela Ursache, Mihaela Turturică, Gabriela Râpeanu, Nicoleta Stănciuc
Sea buckthorn has gained importance as a versatile nutraceutical, due to its high nutritive value in terms of carotenoids content. β-Lactoglobulin (β-LG) is a natural carrier for various bioactive compounds. In this study, the effect of thermal treatment in the temperature range of 25 to 100°C for 15min on the complex formed by β-LG and carotenoids from sea buckthorn was reported, based on fluorescence spectroscopy, molecular docking and molecular dynamics simulation results. Also, the berries extracts were analyzed for their carotenoids content...
October 17, 2016: Spectrochimica Acta. Part A, Molecular and Biomolecular Spectroscopy
Jing Li, Aping Niu, Chun-Jiao Lu, Jing-Hui Zhang, Muhammad Junaid, Phyllis R Strauss, Ping Xiao, Xiao Wang, Yi-Wei Ren, De-Sheng Pei
Landfill leachate (LL) is harmful to aquatic environment because it contains high concentrations of dissolved organic matter, inorganic components, heavy metals, and other xenobiotics. Thus, the remediation of LL is crucial for environmental conservation. Here, a potential application of the forward osmosis (FO) filtration process with ammonium bicarbonate (NH4HCO3) as a draw solution (DS) was investigated to remediate membrane bioreactor-treated LL (M-LL). After the leachate treatment, the toxicity and removal efficiencies of polycyclic aromatic hydrocarbons (PAHs) were evaluated using zebrafish and cultured human cells...
October 21, 2016: Chemosphere
Zahra Aghili, Saba Taheri, Hojjat Alizadeh Zeinabad, Leila Pishkar, Ali Akbar Saboury, Arash Rahimi, Mojtaba Falahati
Herein, the interaction of hen egg white lysozyme (HEWL) with iron nanoparticle (Fe NP) was investigated by spectroscopic and docking studies. The zeta potential analysis revealed that addition of Fe NP (6.45±1.03 mV) to HEWL (8.57±0.54 mV) can cause to greater charge distribution of nanoparticle-protein system (17.33±1.84 mV). In addition, dynamic light scattering (DLS) study revealed that addition of Fe NP (92.95±6.11 nm) to HEWL (2.68±0.37 nm) increases suspension potential of protein/nanoparticle system (51...
2016: PloS One
Sunil K George, Mehran Abolbashari, John D Jackson, Tamer Aboushwareb, Anthony Atala, James J Yoo
Chronic kidney disease (CKD) occurs when certain conditions cause the kidneys to gradually lose function. For patients with CKD, renal transplantation is the only treatment option that restores kidney function. In this study, we evaluated primary renal cells obtained from diseased kidneys to determine whether their normal phenotypic and functional characteristics are retained, and could be used for cell therapy. Primary renal cells isolated from both normal kidneys (NK) and diseased kidneys (CKD) showed similar phenotypic characteristics and growth kinetics...
2016: PloS One
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