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Proximity ligation assay

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https://www.readbyqxmd.com/read/28920103/existence-of-brain-5-ht1a-5-ht2a-isoreceptor-complexes-with-antagonistic-allosteric-receptor-receptor-interactions-regulating-5-ht1a-receptor-recognition
#1
Dasiel O Borroto-Escuela, Xiang Li, Alexander O Tarakanov, David Savelli, Manuel Narváez, Kirill Shumilov, Yuniesky Andrade-Talavera, Antonio Jimenez-Beristain, Bartosz Pomierny, Zaida Díaz-Cabiale, Riccardo Cuppini, Patrizia Ambrogini, Maria Lindskog, Kjell Fuxe
Studies on serotonin-selective reuptake inhibitors have established that disturbances in the ascending 5-HT neuron systems and their 5-HT receptor subtypes and collateral networks to the forebrain contribute to the etiology of major depression and are targets for treatment. The therapeutic action of serotonin-selective reuptake inhibitors is of proven effectiveness, but the mechanisms underlying their effect are still unclear. There are many 5-HT subtypes involved; some need to be blocked (e.g., 5-HT2A, 5-HT3, and 5-HT7), whereas others need to be activated (e...
August 31, 2017: ACS Omega
https://www.readbyqxmd.com/read/28910568/network-of-phosphatases-and-hdac-complexes-at-repressed-chromatin
#2
I J de Castro, H A Amin, V Vinciotti, P Vagnarelli
Tight regulation of gene expression is achieved by a variety of protein complexes that selectively bind chromatin, modify it and change its transcription competency. Histone acetylases (HATs) and deacetylases (HDACs) play an important role in this process. They can generate transcriptionally active or inactive chromatin through the addition (HATs) or removal (HDACs) of acetyl groups on histones, respectively. Repo-Man is a Protein Phosphatase 1 targeting subunit that accumulates on chromosomes during mitotic exit and mediates the removal of mitotic histone H3 phosphorylations...
September 14, 2017: Cell Cycle
https://www.readbyqxmd.com/read/28883043/ffar2-ffar3-receptor-heteromerization-modulates-short-chain-fatty-acid-sensing
#3
Zhiwei Ang, Ding Xiong, Min Wu, Jeak Ling Ding
Free fatty acid receptors (FFAR2/FFA2/GPR43 and FFAR3/FFA3/GPR41) are mammalian receptors for gut microbiota-derived short-chain fatty acids (SCFAs). These receptors are promising drug targets for obesity, colitis, colon cancer, asthma, and arthritis. Here, we demonstrate that FFAR2 and FFAR3 interact to form a heteromer in primary human monocytes and macrophages via proximity ligation assay, and during heterologous expression in HEK293 cells via bimolecular fluorescence complementation and fluorescence resonance energy transfer...
September 7, 2017: FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology
https://www.readbyqxmd.com/read/28882814/opsin-3-and-4-mediate-light-induced-pulmonary-vasorelaxation-that-is-potentiated-by-g-protein-receptor-kinase-2-inhibition
#4
Sebastian Barreto Ortiz, Daijiro Hori, Yohei Nomura, Xin Yun, Haiyang Jiang, Hwanmee Yong, James Chen, Sam Paek, Deepesh Pandey, Gautam Sikka, Anil Bhatta, Andrew Gillard, Jochen Steppan, Jae Hyung Kim, Hideo Adachi, Viachaslau Barodka, Lewis Romer, Steven S An, Larissa A Shimoda, Lakshmi Santhanam, Dan E Berkowitz
We recently demonstrated that blue light induces vasorelaxation in the systemic mouse circulation, a phenomenon mediated by the non-visual G protein-coupled receptor (GPCR) melanopsin (opsin 4; Opn4). Here we tested the hypothesis that non-visual opsins mediate photorelaxation in the pulmonary circulation. We discovered Opsin 3 (Opn3), Opn4, and G-protein coupled receptor kinase 2 (GRK2) in rat pulmonary arteries (PAs) and in pulmonary arterial smooth muscle cells (PASMCs), where the opsins interact directly with GRK2 as demonstrated with a proximity ligation assay...
September 7, 2017: American Journal of Physiology. Lung Cellular and Molecular Physiology
https://www.readbyqxmd.com/read/28881621/connexin40-controls-endothelial-activation-by-dampening-nf%C3%AE%C2%BAb-activation
#5
Jean-Francois Denis, K E Ludwig Scheckenbach, Anna Pfenniger, Merlijn J Meens, Rob Krams, Lucile Miquerol, Steven Taffet, Marc Chanson, Mario Delmar, Brenda R Kwak
Connexins are proteins forming gap junction channels for intercellular communication. Connexin40 (Cx40) is highly expressed by endothelial cells (ECs) of healthy arteries but this expression is lost in ECs overlying atherosclerotic plaques. Low/oscillatory shear stress observed in bends and bifurcations of arteries is atherogenic partly through activation of the pro-inflammatory NFκB pathway in ECs. In this study, we investigated the relation between shear stress, Cx40 and NFκB. Shear stress-modifying casts were placed around carotid arteries of mice expressing eGFP under the Cx40 promoter (Cx40(+/eGFP) )...
August 1, 2017: Oncotarget
https://www.readbyqxmd.com/read/28878211/assay-to-visualize-specific-protein-oxidation-reveals-spatio-temporal-regulation-of-shp2
#6
Ryouhei Tsutsumi, Jana Harizanova, Rabea Stockert, Katrin Schröder, Philippe I H Bastiaens, Benjamin G Neel
Reactive oxygen species are produced transiently in response to cell stimuli, and function as second messengers that oxidize target proteins. Protein-tyrosine phosphatases are important reactive oxygen species targets, whose oxidation results in rapid, reversible, catalytic inactivation. Despite increasing evidence for the importance of protein-tyrosine phosphatase oxidation in signal transduction, the cell biological details of reactive oxygen species-catalyzed protein-tyrosine phosphatase inactivation have remained largely unclear, due to our inability to visualize protein-tyrosine phosphatase oxidation in cells...
September 6, 2017: Nature Communications
https://www.readbyqxmd.com/read/28878041/characteristics-of-29-novel-atypical-solute-carriers-of-major-facilitator-superfamily-type-evolutionary-conservation-predicted-structure-and-neuronal-co-expression
#7
Emelie Perland, Sonchita Bagchi, Axel Klaesson, Robert Fredriksson
Solute carriers (SLCs) are vital as they are responsible for a major part of the molecular transport over lipid bilayers. At present, there are 430 identified SLCs, of which 28 are called atypical SLCs of major facilitator superfamily (MFS) type. These are MFSD1, 2A, 2B, 3, 4A, 4B, 5, 6, 6 L, 7, 8, 9, 10, 11, 12, 13A, 14A and 14B; SV2A, SV2B and SV2C; SVOP and SVOPL; SPNS1, SPNS2 and SPNS3; and UNC93A and UNC93B1. We studied their fundamental properties, and we also included CLN3, an atypical SLC not yet belonging to any protein family (Pfam) clan, because its involvement in the same neuronal degenerative disorders as MFSD8...
September 2017: Open Biology
https://www.readbyqxmd.com/read/28867591/heteromeric-trpv4-trpc1-channels-mediate-calcium-sensing-receptor-induced-nitric-oxide-production-and-vasorelaxation-in-rabbit-mesenteric-arteries
#8
Harry Z E Greenberg, Simonette R E Carlton-Carew, Dhanak M Khan, Alexander K Zargaran, Kazi S Jahan, W-S Vanessa Ho, Anthony P Albert
Stimulation of calcium-sensing receptors (CaSR) by increasing the external calcium concentration (Ca(2+)]o) induces endothelium-dependent vasorelaxation through nitric oxide (NO) production and activation of intermediate Ca(2+)-activated K(+) currents (IKCa) channels in rabbit mesenteric arteries. The present study investigates the potential role of heteromeric TRPV4-TRPC1 channels in mediating these CaSR-induced vascular responses. Immunocytochemical and proximity ligation assays showed that TRPV4 and TRPC1 proteins were expressed and co-localised at the plasma membrane of freshly isolated endothelial cells (ECs)...
September 2017: Vascular Pharmacology
https://www.readbyqxmd.com/read/28864210/inhibition-of-the-mevalonate-pathway-by-simvastatin-interferes-with-mast-cell-degranulation-by-disrupting-the-interaction-between-rab27a-and-double-c2-alpha-proteins
#9
Muhammad Novrizal Abdi Sahid, Shuang Liu, Takeshi Kiyoi, Kazutaka Maeyama
Statins are well-known inhibitors of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase, which block the mevalonate pathway. The activity of statins not only decreases cholesterol levels but also ameliorates inflammation and modulates the immune system. In this study, we investigated the effects of simvastatin on histamine release using rat basophilic leukaemia (RBL-2H3) cells, and examined its interaction with proteins involved in the exocytosis process. Treatment with simvastatin for 24h inhibited histamine release in RBL-2H3 cells in a concentration-dependent manner after stimulation with dinitrophenylated bovine serum albumin (DNP-BSA, as an antigen), ionomycin (a calcium ion [Ca(2+)] ionophore), and thapsigargin (an inhibitor of Ca(2+)-ATPase in the endoplasmic reticulum)...
August 29, 2017: European Journal of Pharmacology
https://www.readbyqxmd.com/read/28862946/%C3%AE-1-adrenergic-receptors-function-within-hetero-oligomeric-complexes-with-atypical-chemokine-receptor-3-and-chemokine-c-x-c-motif-receptor-4-in-vascular-smooth-muscle-cells
#10
Lauren J Albee, Jonathan M Eby, Abhishek Tripathi, Heather M LaPorte, Xianlong Gao, Brian F Volkman, Vadim Gaponenko, Matthias Majetschak
BACKGROUND: Recently, we provided evidence that α1-adrenergic receptors (ARs) in vascular smooth muscle are regulated by chemokine (C-X-C motif) receptor (CXCR) 4 and atypical chemokine receptor 3 (ACKR3). While we showed that CXCR4 controls α1-ARs through formation of heteromeric receptor complexes in human vascular smooth muscle cells (hVSMCs), the molecular basis underlying cross-talk between ACKR3 and α1-ARs is unknown. METHODS AND RESULTS: We show that ACKR3 agonists inhibit inositol trisphosphate production in hVSMCs on stimulation with phenylephrine...
August 17, 2017: Journal of the American Heart Association
https://www.readbyqxmd.com/read/28855353/met-grb2-signaling-associated-complexes-correlate-with-oncogenic-met-signaling-and-sensitivity-to-met-kinase-inhibitors
#11
Matthew A Smith, Thomas Licata, Aliya Lakhani, Marileila Varella-Garcia, Hans-Ulrich Schildhaus, Vincent Vuaroqueaux, Balazs Halmos, Alain Borczuk, Y Ann Chen, Ben C Creelan, Theresa A Boyle, Eric B Haura
PURPOSE: Targeting MET in cancer is hampered by lack of diagnostics that accurately reflect high MET signaling and dependence. We hypothesized that assays reflecting MET signaling associated protein complexes could redefine tumors dependent on MET and could add additional precision beyond genomic assessments. EXPERIMENTAL DESIGN: We utilized biochemical approaches, cellular viability studies and proximity ligation assays to assess MET dependence. We examined MET signaling complexes in lung cancer patient specimens (N=406) and patient-derived xenograft models of solid tumors (N=308)...
August 29, 2017: Clinical Cancer Research: An Official Journal of the American Association for Cancer Research
https://www.readbyqxmd.com/read/28842709/the-parkinson-s-disease-associated-gpr37-receptor-interacts-with-striatal-adenosine-a2a-receptor-controlling-its-cell-surface-expression-and-function-in-vivo
#12
Xavier Morató, Rafael Luján, Marc López-Cano, Jorge Gandía, Igor Stagljar, Masahiko Watanabe, Rodrigo A Cunha, Víctor Fernández-Dueñas, Francisco Ciruela
G protein-coupled receptor 37 (GPR37) is an orphan receptor associated to Parkinson's disease (PD) neuropathology. Here, we identified GPR37 as an inhibitor of adenosine A2A receptor (A2AR) cell surface expression and function in vivo. In addition, we showed that GPR37 and A2AR do oligomerize in the striatum. Thus, a close proximity of GPR37 and A2AR at the postsynaptic level of striatal synapses was observed by double-labelling post-embedding immunogold detection. Indeed, the direct receptor-receptor interaction was further substantiated by proximity ligation in situ assay...
August 25, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28783179/reciprocal-antagonism-between-the-netrin-1-receptor-uncoordinated-phenotype-5a-unc5a-and-the-hepatitis-c-virus
#13
M-L Plissonnier, T Lahlali, M Raab, M Michelet, C Romero-López, M Rivoire, K Strebhardt, D Durantel, M Levrero, P Mehlen, F Zoulim, R Parent
Hepatitis C virus (HCV) infection is a leading cause of hepatocellular carcinoma (HCC), mainly through cirrhosis induction, spurring research for a deeper understanding of HCV versus host interactions in cirrhosis. The present study investigated crosstalks between HCV infection and UNC5A, a netrin-1 dependence receptor that is inactivated in cancer. UNC5A and HCV parameters were monitored in patients samples (n=550) as well as in in vitro. In patients, UNC5A mRNA expression is significantly decreased in clinical HCV(+) specimens irrespective of the viral genotype, but not in (HBV)(+) liver biopsies, as compared to uninfected samples...
August 7, 2017: Oncogene
https://www.readbyqxmd.com/read/28782625/shifted-golgi-targeting-of-glycosyltransferases-and-%C3%AE-mannosidase-ia-from-giantin-to-gm130-grasp65-results-in-formation-of-high-mannose-n-glycans-in-aggressive-prostate-cancer-cells
#14
Ganapati Bhat, Vishwanath-Reddy Hothpet, Ming-Fong Lin, Pi-Wan Cheng
BACKGROUND: There is a pressing need for biomarkers that can distinguish indolent from aggressive prostate cancer to prevent over-treatment of patients with indolent tumor. METHODS: Golgi targeting of glycosyltransferases was characterized by confocal microscopy after knockdown of GM130, giantin, or both. N-glycans on a trans-Golgi enzyme β4galactosyltransferase-1 isolated by immunoprecipitation from androgen-sensitive and independent prostate cancer cells were determined by matrix-assisted laser desorption-time of flight-mass spectrometry...
August 4, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28771231/fgfr1-is-critical-for-the-anti-endothelial-mesenchymal-transition-effect-of-n-acetyl-seryl-aspartyl-lysyl-proline-via-induction-of-the-map4k4-pathway
#15
Jinpeng Li, Sen Shi, Swayam Prakash Srivastava, Munehiro Kitada, Takako Nagai, Kyoko Nitta, Miyuki Kohno, Keizo Kanasaki, Daisuke Koya
Endothelial-to-mesenchymal transition (EndMT) has been shown to contribute to organ fibrogenesis, and we have reported that the anti-EndMT effect of N-acetyl-seryl-aspartyl-lysyl-proline (AcSDKP) is associated with restoring expression of diabetes-suppressed fibroblast growth factor receptor (FGFR), the key anti-EndMT molecule. FGFR1 is the key inhibitor of EndMT via the suppression of the transforming growth factor β (TGFβ) signaling pathway, and mitogen-activated protein kinase kinase kinase kinase 4 (MAP4K4) inhibits integrin β1, a key factor in activating TGFβ signaling and EndMT...
August 3, 2017: Cell Death & Disease
https://www.readbyqxmd.com/read/28764767/monoamine-oxidase-b-is-elevated-in-alzheimer-disease-neurons-is-associated-with-%C3%AE-secretase-and-regulates-neuronal-amyloid-%C3%AE-peptide-levels
#16
Sophia Schedin-Weiss, Mitsuhiro Inoue, Lenka Hromadkova, Yasuhiro Teranishi, Natsuko Goto Yamamoto, Birgitta Wiehager, Nenad Bogdanovic, Bengt Winblad, Anna Sandebring-Matton, Susanne Frykman, Lars O Tjernberg
BACKGROUND: Increased levels of the pathogenic amyloid β-peptide (Aβ), released from its precursor by the transmembrane protease γ-secretase, are found in Alzheimer disease (AD) brains. Interestingly, monoamine oxidase B (MAO-B) activity is also increased in AD brain, but its role in AD pathogenesis is not known. Recent neuroimaging studies have shown that the increased MAO-B expression in AD brain starts several years before the onset of the disease. Here, we show a potential connection between MAO-B, γ-secretase and Aβ in neurons...
August 1, 2017: Alzheimer's Research & Therapy
https://www.readbyqxmd.com/read/28758625/the-dengue-virus-non-structural-protein-1-ns1-is-secreted-from-infected-mosquito-cells-via-a-non-classical-caveolin-1-dependent-pathway
#17
Ana C Alcalá, Raiza Hernández-Bravo, Fernando Medina, David S Coll, Jose L Zambrano, Rosa M Del Angel, Juan E Ludert
Dengue virus NS1 is a glycoprotein of 46-50 kDa that is conserved among flaviviruses, associates as a dimer to cell membranes and is secreted as a hexamer to the extracellular milieu. Recent evidence showed that NS1 is secreted efficiently from infected mosquito cells. To explore the secretory route of NS1 in mosquito cells, infected cells were treated with brefeldin A (BFA) and methyl-beta-cyclodextrin (MβCD). The results showed that MβCD, but not BFA, significantly reduced the release of NS1. Moreover, silencing the expression of caveolin-1 (CAV1; a key component of the caveolar system that transports cholesterol inside the cell), but not SAR1 (a GTPase that participates in the classical secretory pathway), also results in a significant reduction of the secretion of NS1...
August 2017: Journal of General Virology
https://www.readbyqxmd.com/read/28744553/brag2a-a-guanine-nucleotide-exchange-factor-for-arf6-is-a-component-of-the-dystrophin-associated-glycoprotein-complex-at-the-photoreceptor-terminal
#18
Hiroyuki Sakagami, Osamu Katsumata, Yoshinobu Hara, Toshikuni Sasaoka, Masahiro Fukaya
Purpose: Mutations in genes encoding the dystrophin-associated glycoprotein complex (DGC) can cause muscular dystrophy and disturb synaptic transmission in the photoreceptor ribbon synapse. However, the molecular composition and specific functions of the photoreceptor DGC remain unknown. Brefeldin A-resistant Arf-GEF 2 (BRAG2), also known as IQSEC1, is a guanine nucleotide exchange factor for ADP-ribosylation factor 6 (Arf6), a critical GTPase that regulates endosomal trafficking and actin cytoskeleton remodeling...
July 1, 2017: Investigative Ophthalmology & Visual Science
https://www.readbyqxmd.com/read/28742858/reduction-of-endoplasmic-reticulum-mitochondria-interactions-in-beta-cells-from-patients-with-type-2-diabetes
#19
Charles Thivolet, Guillaume Vial, Romeo Cassel, Jennifer Rieusset, Anne-Marie Madec
Type 2 diabetes develops when beta cells are not able to fulfill insulin needs. The role of the endoplasmic reticulum-mitochondria junction in coordinating the functions of these two organelles throughout the natural history of type 2 diabetes is determinant and may explain the alterations of insulin biosynthesis. Our goal was to study endoplasmic reticulum and mitochondrial interactions in human beta cells from organ donors with type 2 diabetes. Pancreas samples were obtained via the network for pancreatic organ donors with diabetes (nPOD) based on disease status with 12 subjects with type 2 diabetes and 9 non-diabetic controls...
2017: PloS One
https://www.readbyqxmd.com/read/28742097/mrtadfinder-a-network-modularity-based-approach-to-identify-topologically-associating-domains-in-multiple-resolutions
#20
Koon-Kiu Yan, Shaoke Lou, Mark Gerstein
Genome-wide proximity ligation based assays such as Hi-C have revealed that eukaryotic genomes are organized into structural units called topologically associating domains (TADs). From a visual examination of the chromosomal contact map, however, it is clear that the organization of the domains is not simple or obvious. Instead, TADs exhibit various length scales and, in many cases, a nested arrangement. Here, by exploiting the resemblance between TADs in a chromosomal contact map and densely connected modules in a network, we formulate TAD identification as a network optimization problem and propose an algorithm, MrTADFinder, to identify TADs from intra-chromosomal contact maps...
July 2017: PLoS Computational Biology
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