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https://www.readbyqxmd.com/read/28627886/experimentally-validated-model-enables-debottlenecking-of-in-vitro-protein-synthesis-and-identifies-a-control-shift-under-in-vivo-conditions
#1
Alexander Nieß, Jurek Failmezger, Maike Kuschel, Martin Siemann-Herzberg, Ralf Takors
Cell-free (in vitro) protein systems (CFPS) provide a versatile tool that can be used to investigate different aspects of the transcription-translation machinery by reducing cells to the basic functions of protein formation. Recent improvements in reaction stability and lysate preparation offer the potential to expand the scope of in vitro biosynthesis from a research tool to a multifunctional and versatile platform for protein production and synthetic biology. To date, even the best-performing CFPS are drastically slower than in vivo references...
June 19, 2017: ACS Synthetic Biology
https://www.readbyqxmd.com/read/28627652/eriodictyol-7%C3%A2-o%C3%A2-%C3%AE-%C3%A2-d-glucopyranoside-from-coreopsis%C3%A2-tinctoria-nutt-ameliorates-lipid-disorders-via-protecting-mitochondrial-function-and-suppressing-lipogenesis
#2
Yuyan Liang, Hai Niu, Limei Ma, Dan Du, Li Wen, Qing Xia, Wen Huang
Coreopsis tinctoria (snow chrysanthemum) has been reported to exert antihyperlipidemic effects. The present study aimed to identify the active compounds of Coreopsis tinctoria and to investigate the molecular mechanisms underlying its effects on lipid dysregulation by measuring lipid levels, reactive oxygen species, lipid peroxidation and fatty acid synthesis. The present results demonstrated that snow chrysanthemum aqueous extracts significantly reduced serum lipid levels and oxidative stress in vivo. The main compounds that were isolated were identified as flavanomarein (compound 1) and eriodictyol 7‑O‑β‑D glucopyranoside (compound 2)...
June 9, 2017: Molecular Medicine Reports
https://www.readbyqxmd.com/read/28625739/changes-in-the-fine-root-proteome-of-fagus-sylvatica-l-trees-associated-with-p-deficiency-and-amelioration-of-p-deficiency
#3
Christoph-Martin Geilfus, Sebastien Christian Carpentier, Aljoša Zavišić, Andrea Polle
Phosphorus is often the least available macronutrient in soil. Lack in phosphorus has detrimental effect on growth and biomass production of European Fagus sylvatica L., a major trees species in temperate forests. In contrast to leaf tissues, few studies have examined changes in the root system and no study has ever investigated the proteomic changes affected in beech roots by a lack in available phosphate (P). Here, we studied roots of young Fagus sylvatica L. trees in their native soils from two forests sites with contrasting availability of P: one P rich and P poor soil...
June 15, 2017: Journal of Proteomics
https://www.readbyqxmd.com/read/28624907/dityrosine-administration-induces-dysfunction-of-insulin-secretion-accompanied-by-diminished-thyroid-hormones-t3-function-in-pancreas-of-mice
#4
Yin-Yi Ding, Zhu-Qing Li, Xiang-Rong Cheng, Yu-Mei Ran, Sha-Ji Wu, Yonghui Shi, Guowei Le
Oxidized tyrosine products are commonly found in food with high protein content and have been demonstrated to cause damage of liver and kidney in our previous studies. Dityrosine (Dityr) is a typical oxidized tyrosine product. Due to its structural homology with thyroid hormones T3, we assumed that one of the endocrine systems most likely considered in connection with its disruption by Dityr may be the T3 action. T3 plays important roles in insulin synthesis, and thyroid hormone resistance (RTH) is associated with the impairment of glucose metabolism...
June 17, 2017: Amino Acids
https://www.readbyqxmd.com/read/28624461/emetine-inhibits-replication-of-rna-and-dna-viruses-without-generating-drug-resistant-virus-variants
#5
Nitin Khandelwal, Yogesh Chander, Thachamvally Riyesh, Krishan Dutt Rawat, Pratibha Saini, Chikkahonnaiah Nishanth, Shalini Sharma, Naresh Jindal, Bhupendra N Tripathi, Sanjay Barua, Naveen Kumar
At a noncytotoxic concentration, emetine was found to inhibit replication of DNA viruses [buffalopoxvirus (BPXV) and bovine herpesvirus 1 (BHV-1)] as well as RNA viruses [peste des petits ruminants virus (PPRV) and Newcastle disease virus (NDV)]. Using the time-of-addition and virus step-specific assays, we showed that emetine treatment resulted in reduced synthesis of viral RNA (PPRV and NDV) and DNA (BPXV and BHV-1) as well as inhibiting viral entry (NDV and BHV-1). In addition, emetine treatment also resulted in decreased synthesis of viral proteins...
June 14, 2017: Antiviral Research
https://www.readbyqxmd.com/read/28623260/qualifying-a-eukaryotic-cell-free-system-for-fluorescence-based-gpcr-analyses
#6
Anne Zemella, Solveig Grossmann, Rita Sachse, Andrei Sonnabend, Michael Schaefer, Stefan Kubick
Membrane proteins are key elements in cell-mediated processes. In particular, G protein-coupled receptors (GPCRs) have attracted increasing interest since they affect cellular signaling. Furthermore, mutations in GPCRs can cause acquired and inheritable diseases. Up to date, there still exist a number of GPCRs that has not been structurally and functionally analyzed due to difficulties in cell-based membrane protein production. A promising approach for membrane protein synthesis and analysis has emerged during the last years and is known as cell-free protein synthesis (CFPS)...
June 16, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28617802/oxidative-stress-affects-processing-of-amyloid-precursor-protein-in-vascular-endothelial-cells
#7
Abebe Muche, Thomas Arendt, Reinhard Schliebs
BACKGROUND: Oxidative stress is thought to be a key player in the pathogenesis of neurodegenerative dementia, including Alzheimer's disease (AD). It has been assumed that oxidative stress contributes to the ß-amyloid deposition in cerebral blood vessels. METHODS: In order to prove this hypothesis, we examined the effect of oxidative stress on the processing of amyloid precursor protein (APP) in primary endothelial cells (EC) derived from cerebral cortical tissue of transgenic Tg2576 mice...
2017: PloS One
https://www.readbyqxmd.com/read/28614063/microfluidic-hydrodynamic-focusing-synthesis-of-polymer-lipid-nanoparticles-for-sirna-delivery
#8
Xueqin Huang, Robert J Lee, Yuhang Qi, Yujing Li, Jiahui Lu, Qingfan Meng, Lesheng Teng, Jing Xie
Small interfering RNAs (siRNAs) are promising as therapeutics for intractable diseases such as cancer. However, efficient and safe delivery of siRNAs in vivo remains a challenge. Polymer-lipid hybrid nanoparticles (P/LNPs) have been evaluated for therapeutic delivery of siRNA. In this study, a microfluidic hydrodynamic focusing (MF) system was used to prepare P/LNPs loaded with VEGF siRNA. P/LNPs made by MF were smaller in particle size and had narrower size distribution compared to P/LNPs formed by bulk mixing (BM)...
May 30, 2017: Oncotarget
https://www.readbyqxmd.com/read/28611809/proteomic-analysis-of-lipid-droplets-from-arabidopsis-aging-leaves-brings-new-insight-into-their-biogenesis-and-functions
#9
Lysiane Brocard, Françoise Immel, Denis Coulon, Nicolas Esnay, Karine Tuphile, Stéphanie Pascal, Stéphane Claverol, Laëtitia Fouillen, Jean-Jacques Bessoule, Claire Bréhélin
Lipid droplets (LDs) are cell compartments specialized for oil storage. Although their role and biogenesis are relatively well documented in seeds, little is known about their composition, structure and function in senescing leaves where they also accumulate. Here, we used a label free quantitative mass spectrometry approach to define the LD proteome of aging Arabidopsis leaves. We found that its composition is highly different from that of seed/cotyledon and identified 28 proteins including 9 enzymes of the secondary metabolism pathways involved in plant defense response...
2017: Frontiers in Plant Science
https://www.readbyqxmd.com/read/28611576/maternal-different-degrees-of-iodine-deficiency-during-pregnant-and-lactation-impair-the-development-of-cerebellar-pinceau-in-offspring
#10
Jing Dong, Heling Song, Yuan Wang, Min Li, Ye Yu, Yi Wang, Jie Chen
Aims: Iodine is critical for synthesis of thyroid hormones (TH). And iodine deficiency (ID) is one of the most significant reasons of intellectual disability and motor memory impairment, although the potential mechanisms are still under investigation. Presently, mild ID and marginal ID are largely ignored problems for women of child bearing age. Mild ID is a subtle form of TH deficiency, which shows low levels of free thyroxine (FT4) and relatively normal free triiodothyronine (FT3) or thyroid stimulation hormone (TSH)...
2017: Frontiers in Neuroscience
https://www.readbyqxmd.com/read/28611396/coupling-neutron-reflectivity-with-cell-free-protein-synthesis-to-probe-membrane-protein-structure-in-supported-bilayers
#11
Thomas Soranzo, Donald K Martin, Jean-Luc Lenormand, Erik B Watkins
The structure of the p7 viroporin, an oligomeric membrane protein ion channel involved in the assembly and release of the hepatitis C virus, was determined from proteins expressed and inserted directly into supported model lipid membranes using cell-free protein expression. Cell-free protein expression allowed (i ) high protein concentration in the membrane, (ii ) control of the protein's isotopic constitution, and (iii ) control over the lipid environment available to the protein. Here, we used cell-free protein synthesis to directly incorporate the hepatitis C virus (HCV) p7 protein into supported lipid bilayers formed from physiologically relevant lipids (POPC or asolectin) for both direct structural measurements using neutron reflectivity (NR) and conductance measurements using electrical impedance spectroscopy (EIS)...
June 13, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28608415/crystal-structural-characterization-reveals-novel-oligomeric-interactions-of-human-voltage-dependent-anion-channel-1
#12
Toshiaki Hosaka, Masateru Okazaki, Tomomi Kimura-Someya, Yoshiko Ishizuka-Katsura, Kaori Ito, Shigeyuki Yokoyama, Kosuke Dodo, Mikiko Sodeoka, Mikako Shirouzu
Voltage-dependent anion channel 1 (VDAC1), which is located in the outer mitochondrial membrane, plays important roles in various cellular processes. For example, oligomerization of VDAC1 is involved in the release of cytochrome c to the cytoplasm, leading to apoptosis. However, it is unknown how VDAC1 oligomerization occurs in the membrane. In the present study, we determined high-resolution crystal structures of oligomeric human VDAC1 (hVDAC1) prepared by using an Escherichia coli cell-free protein synthesis system, which avoided the need for denaturation and refolding of the protein...
June 12, 2017: Protein Science: a Publication of the Protein Society
https://www.readbyqxmd.com/read/28604122/blood-stage-malaria-vaccines-post-genome-strategies-for-the-identification-of-novel-vaccine-candidates
#13
Edward H Ntege, Eizo Takashima, Masayuki Morita, Hikaru Nagaoka, Tomoko Ishino, Takafumi Tsuboi
An efficacious malaria vaccine is necessary to advance the current control measures towards malaria elimination. To-date, only RTS,S/AS01, a leading pre-erythrocytic stage vaccine completed phase 3 trials, but with an efficacy of 28-36% in children, and 18-26% in infants, that waned over time. Blood-stage malaria vaccines protect against disease, and are considered effective targets for the logical design of next generation vaccines to improve the RTS,S field efficacy. Therefore, novel blood-stage vaccine candidate discovery efforts are critical, albeit with several challenges including, high polymorphisms in vaccine antigens, poor understanding of targets of naturally protective immunity, and difficulties in the expression of high AT-rich plasmodial proteins...
June 19, 2017: Expert Review of Vaccines
https://www.readbyqxmd.com/read/28601037/enterococcus-species-for-the-one-pot-biofabrication-of-gold-nanoparticles-characterization-and-nanobiotechnological-applications
#14
Iyabo Christianah Oladipo, Agbaje Lateef, Joseph Adetunji Elegbede, Musibau Adewuyi Azeez, Tesleem Babatunde Asafa, Taofeek Akangbe Yekeen, Akeem Akinboro, Evariste Bosco Gueguim-Kana, Lorika Selomi Beukes, Tolulope Oluyomi Oluyide, Oluwatoyin Rebecca Atanda
In the current work, cell-free extracts of four strains of non-pathogenic Enterococcus species of food origin, were studied for the green synthesis of gold nanoparticles (AuNPs), and characterized by UV-Vis absorption spectroscopy, Fourier transform infrared spectroscopy (FTIR), and transmission electron microscopy (TEM). The AuNPs were evaluated for their Anopheles gambiae larvicidal, dye degradation, antioxidant and thrombolytic activities. The blue-black colloidal AuNPs which absorbed maximally at 549-552nm were nearly spherical in shape, and crystalline in nature with size of 8-50nm...
June 3, 2017: Journal of Photochemistry and Photobiology. B, Biology
https://www.readbyqxmd.com/read/28599069/plant-mitochondria-synthesize-melatonin-and-enhance-the-tolerance-of-plants-to-drought-stress
#15
Lin Wang, Chao Feng, Xiaodong Zheng, Yan Guo, Fangfang Zhou, Dongqian Shan, Xuan Liu, Jin Kong
Synthesis of melatonin in mitochondria was reported in animals. However, there is no report on whether plant mitochondria also produce melatonin. Herein, we show that plant mitochondria are a major site for melatonin synthesis. In an in vitro study, isolated apple mitochondria had the capacity to generate melatonin. Subcellular localization analysis documented that an apple SNAT isoform, MzSNAT5, was localized in the mitochondria of both Arabidopsis protoplasts and apple callus cells. The kinetic analysis revealed that the recombinant MzSNAT5 protein exhibited high enzymatic activity to catalyze serotonin to N-acetylserotonin with the Km and Vmax of 55 μM and 0...
June 9, 2017: Journal of Pineal Research
https://www.readbyqxmd.com/read/28598022/skin-resistance-to-uvb-induced-oxidative-stress-and-hyperpigmentation-by-the-topical-use-of-lactobacillus-helveticus-ns8-fermented-milk-supernatant
#16
Jingjing Rong, Chuang Shan, Shuzhan Liu, Houfeng Zheng, Chen Liu, Ming Liu, Feng Jin, Li Wang
AIMS: Here, we investigated the preventive properties of the supernatant of Lactobacillus helveticus NS8-fermented milk (NS8-FS) against UV light-induced skin oxidative damage and hyperpigmentation. METHODS AND RESULTS: NS8-FS exhibited significant radical scavenging activity in tests with ABST(+) and DPPH scavenging methods, and as well strongly inhibited 3-morpholinosydnonimine (Sin-1) induced ROS generation in HaCat keratinocytes. Unexpectedly, NS8-FS was found to inhibit melanin production in B16F10 melanoma cells, and to exhibit inhibitory effects both to the enzymatic activity of tyrosinase and the expression of proteins required for melanin synthesis...
June 9, 2017: Journal of Applied Microbiology
https://www.readbyqxmd.com/read/28596531/rf1-attenuation-enables-efficient-non-natural-amino-acid-incorporation-for-production-of-homogeneous-antibody-drug-conjugates
#17
Gang Yin, Heather T Stephenson, Junhao Yang, Xiaofan Li, Stephanie M Armstrong, Tyler H Heibeck, Cuong Tran, Mary Rose Masikat, Sihong Zhou, Ryan L Stafford, Alice Y Yam, John Lee, Alexander R Steiner, Avinash Gill, Kalyani Penta, Sonia Pollitt, Ramesh Baliga, Christopher J Murray, Christopher D Thanos, Leslie M McEvoy, Aaron K Sato, Trevor J Hallam
Amber codon suppression for the insertion of non-natural amino acids (nnAAs) is limited by competition with release factor 1 (RF1). Here we describe the genome engineering of a RF1 mutant strain that enhances suppression efficiency during cell-free protein synthesis, without significantly impacting cell growth during biomass production. Specifically, an out membrane protease (OmpT) cleavage site was engineered into the switch loop of RF1, which enables its conditional inactivation during cell lysis. This facilitates extract production without additional processing steps, resulting in a scaleable extract production process...
June 8, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28595554/sulphur-containing-amino-acids-protective-role-against-free-radicals-and-heavy-metals
#18
Mirjana B Čolović, Vesna M Vasić, Dragan M Djuric, Danijela Z Krstić
Sulphur is an abundant element in biological systems, which plays an important role in processes essential for life as a constituent of proteins, vitamins and other crucial biomolecules. The major source of sulphur for humans is plants being able to use inorganic sulphur in the purpose of sulphur-containing amino acids synthesis. Sulphur-containing amino acids include methionine, cysteine, homocysteine, and taurine. Among them, methionine and cysteine are classified as proteinogenic, canonic amino acids incorporated in proteins structure...
June 8, 2017: Current Medicinal Chemistry
https://www.readbyqxmd.com/read/28593644/efficient-trna-degradation-and-quantification-in-escherichia-coli-cell-extract-using-rnase-coated-magnetic-beads-a-key-step-toward-codon-emancipation
#19
Amin S M Salehi, Mark T Smith, Song-Min Schinn, Jeremy M Hunt, Christina Muhlestein, Joann Diray-Arce, Brent L Nielsen, Bradley C Bundy
Emancipating sense codons toward a minimized genetic code is of significant interest to science and engineering. A key approach toward sense codon emancipation is the targeted in vitro removal of native tRNA. However, challenges remain such as the insufficient depletion of tRNA in lysate-based in vitro systems and the high cost of the purified components system (PURE). Here we used RNase-coated superparamagnetic beads to efficiently degrade E. coli endogenous tRNA. The presented method removes >99% of tRNA in cell lysates, while partially preserving cell-free protein synthesis activity...
June 8, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28587922/molecular-basis-for-k63-linked-ubiquitination-processes-in-double-strand-dna-break-repair-a-focus-on-kinetics-and-dynamics
#20
REVIEW
Brian L Lee, Anamika, J N Mark Glover, Michael J Hendzel, Leo Spyracopoulos
Cells are exposed to thousands of DNA damage events on a daily basis. This damage must be repaired to preserve genetic information, and prevent development of disease. The most deleterious damage is a double strand break (DSB), which is detected and repaired by mechanisms known as non-homologous end joining (NHEJ), and homologous recombination (HR), components of the DNA damage response system. NHEJ is an error prone first line of defense, whereas HR invokes error free repair, and is the focus of this review...
June 3, 2017: Journal of Molecular Biology
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