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Thomas Pleli, Antonia Mondorf, Nerea Ferreiros, Dominique Thomas, Karel Dvorak, Ricardo M Biondi, Dagmar Meyer Zu Heringdorf, Stefan Zeuzem, Gerd Geisslinger, Herbert Zimmermann, Oliver Waidmann, Albrecht Piiper
BACKGROUND/AIMS: Signaling of Gs protein-coupled receptors (GsPCRs) is accomplished by stimulation of adenylyl cyclase, causing an increase of the intracellular cAMP concentration, activation of the intracellular cAMP effectors protein kinase A (PKA) and Epac, and an efflux of cAMP, the function of which is still unclear. METHODS: Activation of adenylyl cyclase by GsPCR agonists or cholera toxin was monitored by measurement of the intracellular cAMP concentration by ELISA, anti-phospho-PKA substrate motif phosphorylation by immunoblotting, and an Epac-FRET assay in the presence and absence of adenosine receptor antagonists or ecto-nucleotide phosphodiesterase/pyrophosphatase2 (eNPP2) inhibitors...
March 27, 2018: Cellular Physiology and Biochemistry
Oranuch Techatharatip, Nunthawan Nowwarote, Suthisa Taebunpakul, Prasit Pavasant
Dental pulp cells release adenosine triphosphate (ATP) in response to intrapulpal pressure and the amount released depends on the magnitude of the pressure. ATP regulates the differentiation of stem cells into adipocytes and osteoblasts. However, it is unknown whether extracellular ATP influences the stemness and osteogenic differentiation of stem cells from human exfoliated deciduous teeth (SHEDs). Therefore, this study investigated the effects of extracellular ATP at a low (0.1 μM) and high (10 μM) concentration on the stemness and osteogenic differentiation of SHEDs...
January 2018: Journal of Cellular Biochemistry
Lin Lyu, Baolu Wang, Chaoyang Xiong, Xiaotian Zhang, Xiaoyan Zhang, Junjie Zhang
Autotaxin (ATX) or ectonucleotide pyrophosphatase/phosphodiesterase 2 (ENPP2) is a secretory glycoprotein and functions as the key enzyme for lysophosphatidic acid generation. The mechanism of ATX protein trafficking is largely unknown. Here, we demonstrated that p23, a member of the p24 protein family, was the protein-sorting receptor required for endoplasmic reticulum (ER) export of ATX. A di-phenylalanine (Phe-838/Phe-839) motif in the human ATX C-terminal region was identified as a transport signal essential for the ATX-p23 interaction...
April 28, 2017: Journal of Biological Chemistry
Fengping Jiao, Xiuxiu Wang, Xinling Song, Huijuan Jing, Shangshang Li, Zhenzhen Ren, Zheng Gao, Jianjun Zhang, Le Jia
In present study, a quadratic regression model based on the response surface methodology (RSM) coupled with Box-Behnken design (BBD) was employed to optimize the enzyme-assisted extraction (EAE) process of enzymatic-extractable Pleurotus djamor polysaccharides (EnPPs). By solving the regression equations and analyzing the model graphs, the optimum conditions were at pH 7.36, water to material ratio 56.78, extraction time 44.77min and extraction temperature 35.36°C, respectively. Under these conditions, the experimental yields of EnPPs reached 3...
May 2017: International Journal of Biological Macromolecules
Luca Palazzo, Casey M Daniels, Joanne E Nettleship, Nahid Rahman, Robert Lyle McPherson, Shao-En Ong, Kazuki Kato, Osamu Nureki, Anthony K L Leung, Ivan Ahel
ADP-ribosylation is a conserved post-translational protein modification that plays a role in all major cellular processes, particularly DNA repair, transcription, translation, stress response and cell death. Hence, dysregulation of ADP-ribosylation is linked to the physiopathology of several human diseases including cancers, diabetes and neurodegenerative disorders. Protein ADP-ribosylation can be reversed by the macrodomain-containing proteins PARG, TARG1, MacroD1 and MacroD2, which hydrolyse the ester bond known to link proteins to ADP-ribose as well as consecutive ADP-ribose subunits; targeting this bond can thus result in the complete removal of the protein modification or the conversion of poly(ADP-ribose) to mono(ADP-ribose)...
September 2016: FEBS Journal
Thiago P Chaves, Felipe Hugo A Fernandes, Cleildo P Santana, Jocimar S Santos, Francinalva D Medeiros, Délcio C Felismino, Vanda L Santos, Raïssa Mayer R Catão, Henrique Douglas M Coutinho, Ana Cláudia D Medeiros
Poincianella pyramidalis (Tul.) LP Queiroz (Fabaceae) is an endemic tree of northeastern Brazil, occurring mainly in the Caatinga. Its medicinal use is widespread and is an important therapeutic option against diarrhea, dysentery, and respiratory and urinary infections, among other diseases. In this study we determined the chemical marker and evaluated the interaction between P. pyramidalis extract and a commercial antimicrobial through the use of biological and analytical models. To obtain the extract, an ethanol-water mixture (50:50 v/v) was used as solvent...
2016: PloS One
Ying Sun, Weici Zhang, Jilly F Evans, Annarosa Floreani, Zhengsheng Zou, Yukiko Nishio, Ruizhao Qi, Patrick S C Leung, Christopher L Bowlus, M Eric Gershwin
Autotaxin (ATX) is a 125-kD type II ectonucleotide pyrophosphatase/phosphodiesterase (ENPP2 or NPP2) originally discovered as an unknown "autocrine motility factor" in human melanoma cells. In addition to its pyrophosphatase/phosphodiesterase activities ATX has lysophospholipase D (lysoPLD) activity, catalyzing the conversion of lysophosphatidylcholine (LPC) into lysophosphatidic acid (LPA). ATX is the only ENPP family member with lysoPLD activity and it produces most of the LPA in circulation. In support of this, ATX heterozygous mice have 50% of normal LPA plasma levels...
August 2016: Autoimmunity Reviews
Anwei Cheng, Caijing Han, Xixiu Fang, Jinyue Sun, Xiangyan Chen, Fachun Wan
BACKGROUND: Plant polyphenols are rich in blueberries that have a wide range of properties beneficial to human health. There are two types, according to the solubility of polyphenols, which were defined as extractable polyphenols (EPP) and non-extractable polyphenols (NEPP), respectively. At present, in most of reports, 'total polyphenol' refers only to EPP excluding NEPP. In this paper, the effects of EPP and NEPP on lipopolysaccharides (LPS) induced production of nitric oxide (NO) and gene expression of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) in RAW264...
August 2016: Journal of the Science of Food and Agriculture
Martina Nowak-Machen, Martin Lange, Mark Exley, Sherry Wu, Anny Usheva, Simon C Robson
Hyperoxia is still broadly used in clinical practice in order to assure organ oxygenation in critically ill patients, albeit known toxic effects. In this present study, we hypothesize that lysophosphatidic acid (LPA) mediates NKT cell activation in a mouse model of hyperoxic lung injury. In vitro, pulmonary NKT cells were exposed to hyperoxia for 72 h, and the induction of the ectonucleotide pyrophosphatase/phosphodiesterase 2 (ENPP-2) was examined and production of lysophosphatidic acid (LPA) was measured...
December 2015: Purinergic Signalling
Lorenzo Federico, Kang Jin Jeong, Christopher P Vellano, Gordon B Mills
The ectonucleotide pyrophosphatase/phosphodiesterase type 2, more commonly known as autotaxin (ATX), is an ecto-lysophospholipase D encoded by the human ENNP2 gene. ATX is expressed in multiple tissues and participates in numerous key physiologic and pathologic processes, including neural development, obesity, inflammation, and oncogenesis, through the generation of the bioactive lipid, lysophosphatidic acid. Overwhelming evidence indicates that altered ATX activity leads to oncogenesis and cancer progression through the modulation of multiple hallmarks of cancer pathobiology...
January 2016: Journal of Lipid Research
Krzysztof A Tomaszewski, Dariusz Adamek, Artur Pasternak, Roman Głowacki, Romana Tomaszewska, Jerzy A Walocha
The aim of this study was to clarify the relationship between the expression of ALP, ANK, ENPP-1, OPN and TGF-β1 in the intervertebral disc (IVD), and cervical vertebral endplate calcification and degeneration. Sixty cervical IVDs were excised from 30 human cadavers. Each cadaver was assessed macroscopically for degeneration (Thompson's classification), and then underwent histological processing, regular staining (hematoxylin and eosin, Masson-Goldner trichrome and alcian blue-PAS), immunohistochemistry (ALP, ANK, ENPP-1, OPN and TGF-β1), microscopic degeneration grading (Boos classification), and assessment of endplate calcification...
October 2014: Polish Journal of Pathology: Official Journal of the Polish Society of Pathologists
Ole K Greiner-Tollersrud
Ecto-nucleotide phosphodiesterase/pyrophosphatase 6 (eNPP6) is a glycosylphosphatidylinositol (GPI)-anchored alkaline lysophospholipase C which is predominantly expressed in brain myelin and kidney. Due to shedding of the GPI-anchor eNPP6 occurs also as a soluble isoform (s-eNPP6). eNPP 6 consists of two identical monomers of 55 kDa joined by a disulfide bridge, and possesses four N-glycans in each monomer. In brain s-eNPP6 the N-glycans are mainly hybrid and high mannose type structures, reminiscent of processed mannose-6-phosphorylated glycans...
November 2014: Neurochemical Research
Junko Morita, Kazuki Kato, Emiko Mihara, Ryuichiro Ishitani, Junichi Takagi, Hiroshi Nishimasu, Junken Aoki, Osamu Nureki
Enpp (ectonucleotide phosphodiesterase/pyrophosphatase) 6 is a membrane-bound glycoprotein that hydrolyzes choline-containing compounds such as lysophosphatidylcholine and glycerophosphorylcholine, and presumably participates in choline metabolism. The catalytic domain of mouse Enpp6 was expressed in HEK293T cells, purified using the TARGET tag/P20.1-Sepharose system and crystallized. An X-ray diffraction data set was collected to 1.8 Å resolution. The crystal belonged to space group P1, with unit-cell parameters a=63...
June 2014: Acta Crystallographica. Section F, Structural Biology Communications
Hong-guang Xu, Zi-rui Li, Hong Wang, Ping Liu, Sheng-nan Xiang, Chuang-dong Wang, Xiao-ling Zhang
BACKGROUND: Ectonucleotide pyrophosphatase/phosphodiesterase (ENPP)-1 is a membrane-bound protein that catalyzes the hydrolysis of extracellular nucleoside triphosphates to monophosphate and extracellular inorganic pyrophosphate (ePPi). Mechanical stimulation regulates ENPP-1 expression. This study sought to investigate the changes in ENPP-1 expression after stimulation using cyclic mechanical tension (CMT). METHODS: Rat end-plate chondrocytes were cultured and subjected to CMT (at 3%, 6%, and 9% elongation) for 20, 40, and 60 minutes to observe changes in the expression of ENPP-1...
October 2013: Chinese Medical Journal
Hong-guang Xu, Zi-rui Li, Hong Wang, Ping Liu, Sheng-nan Xiang, Chuang-dong Wang, Xiao-ling Zhang
OBJECTIVE: To investigate the relationship between ectonucleotide pyrophosphatase phosphodiesterase-1(ENPP-1) expression and transforming growth factor beta 1 (TGF-β1) of end-plate chondrocytes after stimulation with intermittent cyclic mechanical tension (ICMT) by using an FX-4000T Flexercell Tension Plus unit. METHODS: Rat end-plate chondrocytes were cultured and ICMT (strain at 0.5 Hz sinusoidal curve at 10% elongation) applied for 7 days for 4 h/day and cultured for a further 2 days...
February 2013: Orthopaedic Surgery
Linn Greiner-Tollersrud, Thomas Berg, Hilde M F R Stensland, Gry Evjen, Ole K Greiner-Tollersrud
Glycerophosphocholine choline phosphodiesterase (GPC-Cpde) is a glycosylphosphatidylinositol (GPI)-anchored alkaline hydrolase that is expressed in the brain and kidney. In brain the hydrolase is synthesized by the oligodendrocytes and expressed on the myelin membrane. There are two forms of brain GPC-Cpde, a membrane-linked (mGPC-Cpde) and a soluble (sGPC-Cpde). Here we report the characterisation sGPC-Cpde from bovine brain. The amino acid sequence was identical to ectonucleotide pyrophosphatase/phosphodiesterase 6 (eNPP6) precursor, lacking the N-terminal signal peptide region and a C-terminal stretch, suggesting that the hydrolase was solubilised by C-terminal proteolysis, releasing the GPI-anchor...
February 2013: Neurochemical Research
Jens Hausmann, Anastassis Perrakis, Wouter H Moolenaar
Autotaxin (ATX or ENPP2) is an ectonucleotide pyrophosphatase/phosphodiesterase (ENPP) that functions as a secreted lysophospholipase D to produce the multifunctional lipid mediator lysophosphatidic acid (LPA) from more complex lysophospholipids. LPA acts on distinct G protein-coupled receptors thereby activating multiple signaling cascades and cellular responses. The ATX-LPA signaling axis is implicated in a remarkably wide variety of physiological and pathological processes, ranging from vascular and neural development to lymphocyte homing, fibrosis and cancer...
January 2013: Advances in Biological Regulation
Hong-Guang Xu, Xiao-Ling Zhang, Xiao-Hai Zhang, Hong Wang, Rui-Ping Jia, Wen-Xue Tong, Hai-Jun Tong
OBJECTIVE: To explore the relationship between endogenous transforming growth factor (TGF)-β1 and calcification-related genes through an in vitro degeneration model by propagating rat endplate chondrocytes during a natural degeneration process. METHODS: Endplate chondrocytes were extracted from rat lumbar vertebrae, isolated by enzyme digestion and P2 and P4 generations selected for a 6-day in vitro culture. The specimens were photographed microscopically to observe the cellular differences by alizarin red staining...
August 23, 2011: Zhonghua Yi Xue za Zhi [Chinese medical journal]
B C Eke, T Coban, N Vural, M Iscan
The adult male and female rats were exposed to cigarette smoke (CS) 5 times a day, with 1 h intervals, for 3 days in a chamber where smoke and fresh air lead alternatively and were killed 16 h after the last treatments and hepatic and pulmonary monooxygenase (MO) activities (aniline-4-hydroxylase, AH; aminopyrine-N-demethylase, AMND; 7-ethoxyresorufin-O-deethylase, EROD; p-nitroanisole-O-demethylase, p-NAOD), lipid peroxidation (LP) and reduced glutathione (GSH) levels and glutathione-S-transferases (GSTs) activities toward several substrates (1-chloro-2,4-dinitrobenzene, CDNB; 1,2-dichloro-4-nitrobenzene, DCNB; ethacrynic acid, EAA; 1,2-epoxy-3-(p-nitrophenoxy)-propane, ENPP) were determined...
January 1998: Environmental Toxicology and Pharmacology
Hong-Guang Xu, Chun-Jiang Hu, Hong Wang, Ping Liu, Xiao-Ming Yang, Yu Zhang, Ling-Ting Wang
We investigated the effects of mechanical strain on the progressive ankylosis (ANK) gene and extracellular nucleotide phosphatase/phosphodiesterase (ENPP)1 mRNA expression and TGF-β1 protein expression in rat endplate chondrocytes in vitro. Endplate chondrocytes were isolated and cultured in vitro. Following identification with toluidine blue and immunocytochemical staining, chondrocytes were subjected to 10% elongation with various frequencies (0.5, 1, 1.5 and 2 Hz) using a Flexercell Tension Plus system at various intervals (3, 6, 12, 24, 36 and 48 h)...
September 2011: Molecular Medicine Reports
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