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cell free protein synthesis

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https://www.readbyqxmd.com/read/28733286/the-regulator-ldhr-and-the-d-lactate-dehydrogenase-ldha-of-burkholderia-multivorans-play-a-role-in-carbon-overflow-and-in-planktonic-cellular-aggregates-formation
#1
Inês N Silva, Marcelo J Ramires, Lisa A Azevedo, Ana R Guerreiro, Andreia C Tavares, Jörg D Becker, Leonilde M Moreira
LysR-type transcriptional regulators (LTTR) are the most commonly found regulators in Burkholderia cepacia complex, comprising opportunistic pathogens causing chronic respiratory infections in cystic fibrosis (CF) patients. Despite LTTRs being global regulators of pathogenicity in several bacteria, few have been characterized in Burkholderia Here, we showed that gene ldhR of B. multivorans encoding a LTTR is co-transcribed with ldhA encoding a d-lactate dehydrogenase, and evaluate their implication in virulence traits like exopolysaccharide (EPS) synthesis and biofilm formation...
July 21, 2017: Applied and Environmental Microbiology
https://www.readbyqxmd.com/read/28727010/effect-of-experimentally-increased-protein-supply-to-postpartum-dairy-cows-on-plasma-protein-synthesis-rumen-tissue-proliferation-and-immune-homeostasis
#2
M Larsen, C M Røntved, P K Theil, M Khatun, C Lauridsen, N B Kristensen
The effect of experimentally increasing the postpartum protein supply on plasma protein synthesis, rumen tissue proliferation, and immune homeostasis was studied using 8 periparturient Holstein cows in a complete randomized design. At calving, cows were assigned to abomasal infusion of water (CTRL) or casein (CAS) in addition to a lactation diet. Casein infusion was gradually decreased from 696 ± 1 g/d at +2 d relative to calving (DRTC) to 212 ± 10 g/d at +29 DRTC to avoid excessive supply. Synthesis rate of plasma proteins was measured at -14, +4, +15, and +29 DRTC by measuring [C]Phe isotopic enrichment in arterial plasma free Phe, total plasma proteins, and albumin after 3, 5, and 7 h of jugular ring[C]Phe infusion...
May 2017: Journal of Animal Science
https://www.readbyqxmd.com/read/28726576/recent-developments-in-thiolated-polymeric-hydrogels-for-tissue-engineering-applications
#3
Mani Gajendiran, Jae-Sung Rhee, Kyobum Kim
This review focuses on the recent strategy in the preparation of thiolated polymers and fabrication of their hydrogel matrices. The mechanism involved in the synthesis of thiolated polymers and fabrication of thiolated polymer hydrogels is exemplified with suitable schematic representations reported in the recent literature. The 2-iminothiolane namely "Traut's reagent" has been widely employed for effectively thiolating the natural polymers such as collagen and gelatin which contain free amino group in their back bone...
July 20, 2017: Tissue Engineering. Part B, Reviews
https://www.readbyqxmd.com/read/28707580/doxorubicin-loaded-micelle-targeting-muc1-a-potential-therapeutics-for-triple-negative-breast-cancer-treatment
#4
Supang Khondee, Chuda Chittasupho, Singkome Tima, Songyot Anuchapreeda
BACKGROUND: Triple negative breast cancer (TNBC) is an aggressive disease associated with poor prognosis and lack of validated targeted therapy. Thus chemotherapy is a main adjuvant treatment for TNBC patients, but it associates with severe toxicities. For a better treatment outcome, we developed an alternative therapeutic, doxorubicin (DOX)-loaded micelles targeting human mucin1 protein (MUC1) that is less toxic, more effective and targeted to TNBC. METHODS: From many candidate peptides, QNDRHPR-GGGSK (QND) and HSQLPQV-GGGSK (HSQ), were identified computationally, synthesized and purified using solid phase peptide synthesis and semi-preparative HPLC...
July 12, 2017: Current Drug Delivery
https://www.readbyqxmd.com/read/28706572/a-microwell-printing-fabrication-strategy-for-the-on-chip-templated-biosynthesis-of-protein-microarrays-for-surface-plasmon-resonance-imaging
#5
Gerald Manuel, Andrej Lupták, Robert M Corn
A two-step templated, ribosomal biosynthesis/printing method for the fabrication of protein microarrays for surface plasmon resonance imaging (SPRI) measurements is demonstrated. In the first step, a sixteen component microarray of proteins is created in microwells by cell free on chip protein synthesis; each microwell contains both an in vitro transcription and translation (IVTT) solution and 350 femtomoles of a specific DNA template sequence that together are used to create approximately 40 picomoles of a specific hexahistidine-tagged protein...
September 22, 2016: Journal of Physical Chemistry. C, Nanomaterials and Interfaces
https://www.readbyqxmd.com/read/28704726/bnip3-induction-by-hypoxia-stimulates-fasn-dependent-free-fatty-acid-production-enhancing-therapeutic-potential-of-umbilical-cord-blood-derived-human-mesenchymal-stem-cells
#6
Hyun Jik Lee, Young Hyun Jung, Gee Euhn Choi, So Hee Ko, Sei-Jung Lee, Sang Hun Lee, Ho Jae Han
Mitophagy under hypoxia is an important factor for maintaining and regulating stem cell functions. We previously demonstrated that fatty acid synthase (FASN) induced by hypoxia is a critical lipid metabolic factor determining the therapeutic efficacy of umbilical cord blood-derived human mesenchymal stem cells (UCB-hMSCs). Therefore, we investigated the mechanism of a major mitophagy regulator controlling lipid metabolism and therapeutic potential of UCB-hMSCs. This study revealed that Bcl2/adenovirus E1B 19kDa protein-interacting protein 3 (BNIP3)-dependent mitophagy is important for reducing mitochondrial reactive oxygen species accumulation, anti-apoptosis, and migration under hypoxia...
July 4, 2017: Redox Biology
https://www.readbyqxmd.com/read/28702280/dissecting-limiting-factors-of-the-protein-synthesis-using-recombinant-elements-pure-system
#7
Jun Li, Chi Zhang, Poyi Huang, Erkin Kuru, Eliot T C Forster-Benson, Taibo Li, George M Church
Reconstituted cell-free protein synthesis systems such as the Protein synthesis Using Recombinant Elements (PURE) system give high-throughput and controlled access to in vitro protein synthesis. Here we show that compared with the commercial S30 crude extract based RTS 100 E. coli HY system, the PURE system has less mRNA degradation and produces up to ∼6-fold full-length proteins. However the majority of polypeptides PURE produces are partially translated or inactive since the signal from firefly luciferase (Fluc) translated in PURE is only ∼2/3(rd) of that measured using the RTS 100 E...
2017: Translation
https://www.readbyqxmd.com/read/28694500/resistance-exercise-initiates-mechanistic-target-of-rapamycin-mtor-translocation-and-protein-complex-co-localisation-in-human-skeletal-muscle
#8
Zhe Song, Daniel R Moore, Nathan Hodson, Carl Ward, Jessica R Dent, Mary F O'Leary, Andrew M Shaw, D Lee Hamilton, Sovan Sarkar, Yann-Gaël Gangloff, Troy A Hornberger, Lawrence L Spriet, George J Heigenhauser, Andrew Philp
The mechanistic target of rapamycin (mTOR) is a central mediator of protein synthesis in skeletal muscle. We utilized immunofluorescence approaches to study mTOR cellular distribution and protein-protein co-localisation in human skeletal muscle in the basal state as well as immediately, 1 and 3 h after an acute bout of resistance exercise in a fed (FED; 20 g Protein/40 g carbohydrate/1 g fat) or energy-free control (CON) state. mTOR and the lysosomal protein LAMP2 were highly co-localised in basal samples...
July 10, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28694021/recombinant-and-chemo-bio-orthogonal-synthesis-of-liposomal-thrombomodulin-and-its-antithrombotic-activity
#9
Lin Wang, Rui Jiang, Yang Liu, Maosheng Cheng, Qingyu Wu, Xue-Long Sun
Thrombomodulin (TM) is an endothelial cell membrane protein that acts as a major cofactor in the protein C anticoagulant pathway. The EGF-like domains 4-6 of TM (TM456) are essential for PC activation. In this study, we proposed a liposomal recombinant TM conjugate to mimic the membrane TM structure and its anticoagulant activity. First, a DSPE-PEG2000-TM456 was successfully synthesized by site-specific conjugation of azido-TM456 with DSPE-PEG2000-DBCO via copper-free click chemistry quantitatively. Then, liposome-TM456 was fabricated via direct liposome formation with the DSPE-PEG2000-TM456 and other lipids...
July 7, 2017: Journal of Bioscience and Bioengineering
https://www.readbyqxmd.com/read/28690157/cell-free-protein-synthesis-in-micro-compartments-building-a-minimal-cell-from-biobricks
#10
REVIEW
Haiyang Jia, Michael Heymann, Frank Bernhard, Petra Schwille, Lei Kai
The construction of a minimal cell that exhibits the essential characteristics of life is a great challenge in the field of synthetic biology. Assembling a minimal cell requires multidisciplinary expertise from physics, chemistry and biology. Scientists from different backgrounds tend to define the essence of 'life' differently and have thus proposed different artificial cell models possessing one or several essential features of living cells. Using the tools and methods of molecular biology, the bottom-up engineering of a minimal cell appears in reach...
July 6, 2017: New Biotechnology
https://www.readbyqxmd.com/read/28682541/-effect-of-calcitonin-gene-related-peptide-on-mc3t3-e1-osteoblast-apoptosis-and-autophagy-induced-by-serum-starvation
#11
An Yang, Zhang Huiyu, Guo Junfeng, Li Xin, Yang Yang, Zhang Gang, Tan Yinghui
OBJECTIVE: To study the effect of calcitonin gene-related peptide (CGRP) on apoptosis and autophagy of mouse MC3T3-E1 osteoblast and their interaction and to further clarify protective mechanism of CGRP on osteoblasts. METHODS: MC3T3-E1 osteoblasts of mouse were cultured in vitro. Western blot and flow cytometry were used to detect expressions of microtubule-associated protein 1 light chain 3 (LC3) and P62 protein of MC3T3-E1 osteoblasts cultured with serum culture and serum-free (serum starvation) culture...
April 1, 2017: Hua Xi Kou Qiang Yi Xue za Zhi, Huaxi Kouqiang Yixue Zazhi, West China Journal of Stomatology
https://www.readbyqxmd.com/read/28680592/hif-prolyl-hydroxylase-phd3-regulates-translational-machinery-and-glucose-metabolism-in-clear-cell-renal-cell-carcinoma
#12
Petra Miikkulainen, Heidi Högel, Krista Rantanen, Tomi Suomi, Petri Kouvonen, Laura L Elo, Panu M Jaakkola
BACKGROUND: A key feature of clear cell renal cell carcinoma (ccRCC) is the inactivation of the von Hippel-Lindau tumour suppressor protein (pVHL) that leads to the activation of hypoxia-inducible factor (HIF) pathway also in well-oxygenated conditions. Important regulator of HIF-α, prolyl hydroxylase PHD3, is expressed in high amounts in ccRCC. Although several functions and downstream targets for PHD3 in cancer have been suggested, the role of elevated PHD3 expression in ccRCC is not clear...
2017: Cancer & Metabolism
https://www.readbyqxmd.com/read/28680098/quantitative-characterization-of-the-auxin-inducible-degron-a-guide-for-dynamic-protein-depletion-in-single-yeast-cells
#13
Alexandros Papagiannakis, Janeska J de Jonge, Zheng Zhang, Matthias Heinemann
Perturbations are essential for the interrogation of biological systems. The auxin-inducible degron harbors great potential for dynamic protein depletion in yeast. Here, we thoroughly and quantitatively characterize the auxin-inducible degron in single yeast cells. We show that an auxin concentration of 0.25 mM is necessary for fast and uniform protein depletion between single cells, and that in mother cells proteins are depleted faster than their daughters. Although, protein recovery starts immediately after removal of auxin, it takes multiple generations before equilibrium is reached between protein synthesis and dilution, which is when the original protein levels are restored...
July 5, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28667792/phase-i-study-of-salazosulfapyridine-in-combination-with-cisplatin-and-pemetrexed-for-advanced-non-small-cell-lung-cancer
#14
Kohei Otsubo, Kaname Nosaki, Chiyo K Imamura, Hiroaki Ogata, Akitaka Fujita, Shinya Sakata, Fumihiko Hirai, Gouji Toyokawa, Eiji Iwama, Taishi Harada, Takashi Seto, Mitsuhiro Takenoyama, Takeshi Ozeki, Taisei Mushiroda, Mieko Inada, Junji Kishimoto, Kenji Tsuchihashi, Kentaro Suina, Osamu Nagano, Hideyuki Saya, Yoichi Nakanishi, Isamu Okamoto
Splice variant isoforms of CD44 (CD44v) are a marker of cancer stem cells (CSCs) in solid tumors. They stabilize the xCT subunit of the transporter system xc(-) and thereby promote synthesis of the antioxidant glutathione. Salazosulfapyridine (SASP) is an inhibitor of xCT and suppresses the proliferation of CD44v-positive cancer cells. Chemotherapy-naïve patients with advanced nonsquamous non-small cell lung cancer were enrolled in a dose-escalation study (standard 3 + 3 design) of SASP in combination with cisplatin and pemetrexed...
July 1, 2017: Cancer Science
https://www.readbyqxmd.com/read/28667283/the-role-of-the-flid-c-terminal-domain-in-pentamer-formation-and-interaction-with-flit
#15
Hee Jung Kim, Woongjae Yoo, Kyeong Sik Jin, Sangryeol Ryu, Hyung Ho Lee
Flagellar biogenesis is controlled by a negative feedback loop. When FliD was secreted at the late step of flagellar assembly, the FliD-FliT complex disassembled and free FliT bound to the FlhDC complex, a master regulator of flagellar biogenesis, subsequently inhibiting the overall expression of flagellar proteins. In this study, we analyzed the role of the FliD C-terminal domain in pentamer formation and interaction with FliT. Our study showed that the FliD L443R mutant exists as a monomer in solution, indicating that the Leu443 residue of FliD, which contributes to its interaction with FliT, plays a crucial role in the pentameric oligomerization of FliD...
June 30, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28665321/capsaicin-sensitive-sensory-nerves-are-necessary-for-the-protective-effect-of-ghrelin-in-cerulein-induced-acute-pancreatitis-in-rats
#16
Joanna Bonior, Zygmunt Warzecha, Piotr Ceranowicz, Ryszard Gajdosz, Piotr Pierzchalski, Michalina Kot, Anna Leja-Szpak, Katarzyna Nawrot-Porąbka, Paweł Link-Lenczowski, Michał Pędziwiatr, Rafał Olszanecki, Krzysztof Bartuś, Rafał Trąbka, Beata Kuśnierz-Cabala, Artur Dembiński, Jolanta Jaworek
Ghrelin was shown to exhibit protective and therapeutic effect in the gut. Aim of the study was to investigate the role of sensory nerves (SN) in the protective effect of ghrelin in acute pancreatitis (AP). Studies were performed on male Wistar rats or isolated pancreatic acinar cells. After capsaicin deactivation of sensory nerves (CDSN) or treatment with saline, rats were pretreated intraperitoneally with ghrelin or saline. In those rats, AP was induced by cerulein or pancreases were used for isolation of pancreatic acinar cells...
June 30, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/28660957/inverse-electron-demand-diels-alder-reactions-in-chemical-biology
#17
REVIEW
B L Oliveira, Z Guo, G J L Bernardes
The emerging inverse electron demand Diels-Alder (IEDDA) reaction stands out from other bioorthogonal reactions by virtue of its unmatchable kinetics, excellent orthogonality and biocompatibility. With the recent discovery of novel dienophiles and optimal tetrazine coupling partners, attention has now been turned to the use of IEDDA approaches in basic biology, imaging and therapeutics. Here we review this bioorthogonal reaction and its promising applications for live cell and animal studies. We first discuss the key factors that contribute to the fast IEDDA kinetics and describe the most recent advances in the synthesis of tetrazine and dienophile coupling partners...
June 29, 2017: Chemical Society Reviews
https://www.readbyqxmd.com/read/28653367/significance-of-l-carnitine-for-human-health
#18
REVIEW
María M Adeva-Andany, Isabel Calvo-Castro, Carlos Fernández-Fernández, Cristóbal Donapetry-García, Ana María Pedre-Piñeiro
Carnitine acyltransferases catalyze the reversible transfer of acyl groups from acyl-coenzyme A esters to l-carnitine, forming acyl-carnitine esters that may be transported across cell membranes. l-Carnitine is a wáter-soluble compound that humans may obtain both by food ingestion and endogenous synthesis from trimethyl-lysine. Most l-carnitine is intracellular, being present predominantly in liver, skeletal muscle, heart and kidney. The organic cation transporter-2 facilitates l-carnitine uptake inside cells...
August 2017: IUBMB Life
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
#19
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
#20
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)...
August 2017: Molecular Medicine Reports
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