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Molecular & Cellular Proteomics: MCP

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https://www.readbyqxmd.com/read/28087594/a-human-proteome-array-approach-to-identifying-key-host-proteins-targeted-by-toxoplasma-kinase-rop18
#1
Zhaoshou Yang, Yongheng Hou, Taofang Hao, Hee-Sool Rho, Jun Wan, Yizhao Luan, Xin Gao, Jianping Yao, Aihua Pan, Zhi Xie, Jiang Qian, Wanqin Liao, Heng Zhu, Xingwang Zhou
Toxoplasma kinase ROP18 is a key molecule responsible for the virulence of Toxoplasma gondii; however, the mechanisms by which ROP18 exerts parasite virulence via interaction with host proteins remain limited to a small number of identified substrates. To identify a broader array of ROP18 substrates we successfully purified bioactive mature ROP18 and used it to probe a human proteome array. Sixty-eight new putative host targets were identified. Functional annotation analysis suggested that these proteins have a variety of functions including metabolic process, kinase activity and phosphorylation, cell growth, apoptosis and cell death, and immunity, indicating a pleiotropic role of ROP18 kinase...
January 13, 2017: Molecular & Cellular Proteomics: MCP
https://www.readbyqxmd.com/read/28087593/norovirus-mediated-modification-of-the-translational-landscape-via-virus-and-host-induced-cleavage-of-translation-initiation-factors
#2
Edward Emmott, Frederic Sorgeloos, Sarah L Caddy, Surender Vashist, Stanislav Sosnovtsev, Richard Lloyd, Kate Heesom, Nicolas Locker, Ian Goodfellow
Noroviruses produce viral RNAs lacking a 5' cap structure and instead use a virus-encoded VPg protein covalently linked to viral RNA to interact with translation initiation factors and drive viral protein synthesis. Norovirus infection results in the induction of the innate response leading to interferon stimulated gene (ISG) transcription. However the translation of the induced ISG mRNAs is suppressed. A SILAC-based mass spectrometry approach was employed to analyse changes to protein abundance in both whole cell and m7GTP-enriched samples to demonstrate that diminished host mRNA translation correlates with changes to the composition of the eukaryotic initiation factor complex...
January 13, 2017: Molecular & Cellular Proteomics: MCP
https://www.readbyqxmd.com/read/28082515/impact-of-cystinosin-glycosylation-on-protein-stability-by-differential-dynamic-silac
#3
Nathalie Nevo, Lucie Thomas, Cerina Chhuon, Zuzanna Andrzejewska, Joanna Lipecka, François Guillonneau, Anne Bailleux, Aleksander Edelman, Corinne Antignac, Ida Chiara Guerrera
Cystinosis is a rare autosomal recessive lysosomal storage disorder characterized by intralysosomal accumulation of cystine. The causative gene for cystinosis is CTNS, which encodes the protein cystinosin, a lysosomal proton-driven cystine transporter. Over 100 mutations have been reported, leading to varying disease severity, often in correlation with residual cystinosin activity as a transporter and with maintenance of its protein-protein interactions. In this study, we focus on the ΔITILELP mutation, the only mutation reported that sometimes leads to severe forms, inconsistent with its residual transported activity...
January 12, 2017: Molecular & Cellular Proteomics: MCP
https://www.readbyqxmd.com/read/28077445/human-antiviral-protein-ifix-suppresses-viral-gene-expression-during-hsv-1-infection-and-is-counteracted-by-virus-induced-proteasomal-degradation
#4
Marni S Crow, Ileana M Cristea
The interferon inducible protein X (IFIX), a member of the PYHIN family, was recently recognized as an antiviral factor against infection with herpes simplex virus 1 (HSV-1). IFIX binds viral DNA upon infection and promotes antiviral cytokines expression. How IFIX exerts its host defense functions and whether it is inhibited by the virus remains unknown. Here, we integrated live cell microscopy, proteomics, IFIX domain characterization, and molecular virology to investigate IFIX regulation and antiviral functions during HSV-1 infection...
January 11, 2017: Molecular & Cellular Proteomics: MCP
https://www.readbyqxmd.com/read/28077444/protein-interactions-during-the-flavivirus-and-hepacivirus-life-cycle
#5
Gisa Gerold, Janina Bruening, Bettina Weigel, Thomas Pietschmann
Protein-protein interactions govern biological functions in cells, in the extracellular milieu and at the border between cells and extracellular space. Viruses are small intracellular parasites and thus rely on protein interactions to produce progeny inside host cells and to spread from cell to cell. Usage of host proteins by viruses can have severe consequences e.g. apoptosis, metabolic disequilibria or altered cell proliferation and mobility. Understanding protein interactions during virus infection can thus educate us on viral infection and pathogenesis mechanisms...
January 11, 2017: Molecular & Cellular Proteomics: MCP
https://www.readbyqxmd.com/read/28062799/a-two-pronged-binding-mechanism-of-igg-to-the-neonatal-fc-receptor-controls-complex-stability-and-igg-serum-half-life
#6
Pernille Foged Jensen, Angela Schoch, Vincent Larraillet, Maximiliane Hilger, Tilman Schlothauer, Thomas Emrich, Kasper Dyrbert Rand
The success of recombinant monoclonal immunoglobulins (IgG) is rooted in their ability to target distinct antigens with high affinity combined with an extraordinarily long serum half-life, typically around three weeks. The pharmacokinetics of IgGs is intimately linked to the recycling mechanism of the neonatal Fc receptor (FcRn). For long serum half-life of therapeutic IgGs, the highly pH dependent interaction with FcRn needs to be balanced to allow efficient FcRn binding and release at slightly acidic pH and physiological pH, respectively...
January 6, 2017: Molecular & Cellular Proteomics: MCP
https://www.readbyqxmd.com/read/28062798/quantitative-proteome-analysis-of-mouse-liver-lysosomes-provides-evidence-for-mannose-6-phosphate-independent-targeting-mechanisms-of-acid-hydrolases-in-mucolipidosis-ii
#7
Sandra Markmann, Svenja Krambeck, Christopher J Hughes, Mina Mirzaian, Johannes M F G Aerts, Paul Saftig, Michaela Schweizer, Johannes C P Vissers, Thomas Braulke, Markus Damme
The efficient receptor-mediated targeting of soluble lysosomal proteins to lysosomes requires the modification with mannose 6-phosphate (M6P) residues. Although the absence of M6P results in missorting and hypersecretion of lysosomal enzymes in many cells, normal levels of lysosomal enzymes have been reported in liver of patients lacking the M6P-generating phosphotransferase (PT). The identity of lysosomal proteins depending on M6P has not yet been comprehensively analyzed. In this study we purified lysosomes from liver of PT-defective mice and 67 known soluble lysosomal proteins were quantified using an ion mobility-assisted data independent label-free LC-MS approach...
January 6, 2017: Molecular & Cellular Proteomics: MCP
https://www.readbyqxmd.com/read/28062800/novel-allosteric-pathway-of-eg5-regulation-identified-through-multivariate-statistical-analysis-of-hx-ms-ligand-screening-data
#8
Joey G Sheff, Farshad Farshidfar, Oliver F Bathe, Karen Kopciuk, Francesco Gentile, Jack Tuszynski, Khaled Barakat, David C Schriemer
The mitotic kinesin Eg5 is an important target in cancer chemotherapy. A structurally diverse collection of canonical loop L5 inhibitors engage an allosteric pathway that includes elements of its microtubule binding region. However, recent evidence suggests that Eg5 may permit alternative allosteric mechanisms. Terpendole E, a natural-product Eg5 inhibitor, is active against mutants resistant to canonical loop L5 inhibitors and appears to offer a unique mode of inhibition. To investigate the variety of inhibitor responses, the structure-function properties of eighteen kinesin inhibitors were quantified with hydrogen-exchange mass spectrometry (HX-MS), functional analysis and molecular modeling...
January 5, 2017: Molecular & Cellular Proteomics: MCP
https://www.readbyqxmd.com/read/28062797/targeted-proteomics-for-multiplexed-verification-of-markers-of-colorectal-tumorigenesis
#9
Anuli Christiana Uzozie, Nathalie Selevsek, Asa Wahlander, Paolo Nanni, Jonas Grossmann, Achim Weber, Federico Buffoli, Giancarlo Marra
Targeted proteomic methods can accelerate the verification of multiple tumor marker candidates in large series of patient samples. We utilized the targeted approach known as selected / multiple reaction monitoring (SRM/MRM) to verify potential protein markers of colorectal adenoma identified by our group in previous transcriptomic and quantitative shotgun proteomic studies of a large cohort of precancerous colorectal lesions. We developed SRM assays to reproducibly detect and quantify 25 (62.5%) of the 40 selected proteins in an independent series of precancerous and cancerous tissue samples (19 adenoma/normal mucosa pairs; 17 adenocarcinoma/normal mucosa pairs)...
January 4, 2017: Molecular & Cellular Proteomics: MCP
https://www.readbyqxmd.com/read/28062796/integrated-cellular-and-plasma-proteomics-of-contrasting-b-cell-cancers-reveals-common-unique-and-systemic-signatures
#10
Harvey E Johnston, Matthew J Carter, Kerry L Cox, Melanie Dunscombe, Antigoni Manousopoulou, Paul A Townsend, Spiro D Garbis, Mark S Cragg
Approximately 800,000 leukaemia and lymphoma cases are diagnosed worldwide each year. Burkitt's lymphoma (BL) and chronic lymphocytic leukaemia (CLL), are examples of contrasting B-cell cancers; BL is a highly aggressive lymphoid tumour, frequently affecting children, whilst CLL typically presents as an indolent, slow-progressing leukaemia affecting the elderly. The B-cell-specific over-expression of the myc and tcl1 oncogenes in mice induce spontaneous malignancies modelling BL and CLL, respectively. Quantitative mass spectrometry proteomics and isobaric labelling were employed to examine the biology underpinning contrasting Eμ-myc and Eμ-TCL1 B-cell tumours...
January 4, 2017: Molecular & Cellular Proteomics: MCP
https://www.readbyqxmd.com/read/28062795/silac-based-proteomics-of-primary-human-kidney-cells-reveals-a-novel-link-between-male-sex-hormones-and-impaired-energy-metabolism-in-diabetic-kidney-disease
#11
Sergi Clotet, Maria Jose Soler, Marta Riera, Julio Pascual, Fei Fang, Joyce Zhou, Ihor Batruch, Stella K Vasiliou, Apostolos Dimitromanolakis, Clara Barrios, Eleftherios Diamandis, James Scholey, Ana Konvalinka
Male sex predisposes to many kidney diseases. Considering that androgens exert deleterious effects in a variety of cell types within kidney, we hypothesized that dihydrotestosterone (DHT) would alter the biology of the renal tubular cell by inducing changes in the proteome. We employed stable isotope labeling with amino acids (SILAC) in an indirect spike-in fashion to accurately quantify the proteome in DHT- and 17β-estradiol (EST)-treated human proximal tubular epithelial cells (PTEC). Of the 5043 quantified proteins, 76 were differentially regulated...
January 4, 2017: Molecular & Cellular Proteomics: MCP
https://www.readbyqxmd.com/read/28040698/the-prenylated-proteome-of-plasmodium-falciparum-reveals-pathogen-specific-prenylation-activity-and-drug-mechanism-of-action
#12
Jolyn E Gisselberg, Lichao Zhang, Joshua E Elias, Ellen Yeh
Plasmodium parasites contain several unique membrane compartments in which prenylated proteins may play important roles in pathogenesis. Protein prenylation has also been proposed as an antimalarial drug target since farnesyltransferase inhibitors cause potent growth inhibition of blood-stage Plasmodium However, the specific prenylated proteins that mediate antimalarial activity have yet to be identified. Given the potential for new parasite biology and elucidating drug mechanism-of-action, we performed a large-scale identification of the prenylated proteome in blood-stage P...
December 31, 2016: Molecular & Cellular Proteomics: MCP
https://www.readbyqxmd.com/read/28028127/myofibrillar-z-discs-are-a-protein-phosphorylation-hot-spot-with-pkc-%C3%AE-modulating-protein-dynamics
#13
Lena Reimann, Heike Wiese, Yvonne Leber, Anja N Schwäble, Anna L Fricke, Anne Rohland, Bettina Knapp, Christian D Peikert, Friedel Drepper, Peter F M van der Ven, Gerald Radziwill, Dieter O Fürst, Bettina Warscheid
The Z-disc is a protein-rich structure critically important for the development and integrity of myofibrils, which are the contractile organelles of cross-striated muscle cells. We here used mouse C2C12 myoblast which were differentiated into myotubes followed by electrical pulse stimulation (EPS) to generate contracting myotubes comprising mature Z-discs. Using a quantitative proteomics approach, we analyzed changes in the relative abundance of 2,588 proteins, of which 387 were significantly regulated in myoblasts versus myotubes...
December 27, 2016: Molecular & Cellular Proteomics: MCP
https://www.readbyqxmd.com/read/28007913/vegf-promotes-assembly-of-the-p130cas-interactome-to-drive-endothelial-chemotactic-signalling-and-angiogenesis
#14
Ian M Evans, Susan A Kennedy, Ketevan Paliashvili, Tapesh Santra, Maiko Yamaji, Ruth C Lovering, Gary Britton, Paul Frankel, Walter Kolch, Ian C Zachary
p130Cas is a polyvalent adapter protein essential for cardiovascular development, and with a key role in cell movement. In order to identify the pathways by which p130Cas exerts its biological functions in endothelial cells we mapped the p130Cas interactome and its dynamic changes in response to VEGF using high-resolution mass spectrometry and reconstruction of protein interaction (PPI) networks with the aid of multiple PPI databases. VEGF enriched the p130Cas interactome in proteins involved in actin cytoskeletal dynamics and cell movement, including actin-binding proteins, small GTPases and regulators or binders of GTPases...
December 22, 2016: Molecular & Cellular Proteomics: MCP
https://www.readbyqxmd.com/read/27998951/a-proteomic-survey-indicates-sortilin-as-a-secondary-substrate-of-the-er-translocation-inhibitor-cyclotriazadisulfonamide-cada
#15
Victor Van Puyenbroeck, Elisa Claeys, Dominique Schols, Thomas W Bell, Kurt Vermeire
The small molecule CADA was shown to down-modulate the expression of human CD4 in a signal peptide-dependent way, through inhibition of its co-translational translocation across the ER membrane. Previous studies characterizing general glycoprotein levels and the expression of 14 different cell surface receptors showed selectivity of CADA for human CD4. Here, a PowerBlot western array was used as a screen to analyze the proteome of CADA-treated SUP-T1 human CD4+ T lymphocytes. This high-throughput monoclonal antibody panel-based immunoblotting assay of cellular signaling proteins revealed that only a small subset of the 444 detected proteins was differentially expressed after treatment with CADA...
December 20, 2016: Molecular & Cellular Proteomics: MCP
https://www.readbyqxmd.com/read/27965274/rapid-production-of-virus-protein-microarray-using-protein-microarray-fabrication-through-gene-synthesis-pages
#16
Huan Qi, Huiqiong Zhou, Daniel Mark Czajkowsky, Shujuan Guo, Yang Li, Nan Wang, Yi Shi, Lifeng Lin, Jingfang Wang, De Wu, Sheng-Ce Tao
The high genetic variability of RNA viruses is a significant factor limiting the discovery of effective biomarkers, the development of vaccines, and characterizations of the immune response during infection. Protein microarrays have been shown to be a powerful method in biomarker discovery and the identification of novel protein-protein interaction networks, suggesting that this technique could also be very useful in studies of infectious RNA viruses. However, to date, the amount of genetic material required to produce protein arrays, as well as the time- and labor-intensive procedures typically needed, have limited their more widespread application...
December 13, 2016: Molecular & Cellular Proteomics: MCP
https://www.readbyqxmd.com/read/27956708/structural-analysis-of-variable-domain-glycosylation-of-anti-citrullinated-protein-antibodies-in-rheumatoid-arthritis-reveals-the-presence-of-highly-sialylated-glycans
#17
Lise Hafkenscheid, Albert Bondt, Hans U Scherer, Tom W J Huizinga, Manfred Wuhrer, René E M Toes, Yoann Rombouts
Recently, we showed the unexpectedly high abundance of N-linked glycans on the Fab-domain of Anti-Citrullinated Protein Antibodies (ACPA). As N-linked glycans can mediate a variety of biological functions, we now aimed at investigating the structural composition of the Fab-glycans of ACPA-IgG to better understand their mediated biological effects. ACPA-IgG and non-citrulline specific (control) IgG from plasma and/or synovial fluid of nine ACPA positive rheumatoid arthritis patients were affinity purified. The N-linked glycosylation of total, Fc and F(ab)2 fragments, as well as heavy and light chains of ACPA-IgG and control IgG were analyzed by UHPLC and MALDI-TOF mass spectrometry...
December 12, 2016: Molecular & Cellular Proteomics: MCP
https://www.readbyqxmd.com/read/27956707/a-luciferase-fragment-complementation-assay-to-detect-lipid-droplet-associated-protein-protein-interactions
#18
Petra Kolkhof, Michael Werthebach, Anna van de Venn, Gereon Poschmann, Lili Chen, Michael Welte, Kai Stühler, Mathias Beller
A critical challenge for all organisms is to carefully control the amount of lipids they store. An important node for this regulation is the protein coat present at the surface of lipid droplets (LDs), the intracellular organelles dedicated to lipid storage. Only limited aspects of this regulation are understood so far. For the probably best characterized case, the regulation of lipolysis in mammals, some of the major protein players have been identified, and it has been established that this process crucially depends on an orchestrated set of protein-protein interactions...
December 12, 2016: Molecular & Cellular Proteomics: MCP
https://www.readbyqxmd.com/read/27940637/an-optimized-chromatographic-strategy-for-multiplexing-in-parallel-reaction-monitoring-mass-spectrometry-insights-from-quantitation-of-activated-kinases
#19
Anatoly Urisman, Rebecca S Levin, John D Gordan, James T Webber, Hilda Hernandez, Yasushi Ishihama, Kevan M Shokat, Alma L Burlingame
Reliable quantitation of protein abundances in defined sets of cellular proteins is critical to numerous biological applications. Traditional immunodetection-based methods are limited by the quality and availability of specific antibodies, especially for site-specific post-translational modifications. Targeted proteomic methods, including the recently developed parallel reaction monitoring (PRM) mass spectrometry, have enabled accurate quantitative measurements of up to a few hundred specific target peptides...
December 11, 2016: Molecular & Cellular Proteomics: MCP
https://www.readbyqxmd.com/read/27941081/a-multivariate-mixture-model-to-estimate-the-accuracy-of-glycosaminoglycan-identifications-made-by-ms-ms-and-database-search
#20
Yulun Chiu, Paul Schliekelman, Ron Orlando, Joshua S Sharp
We present a statistical model to estimate the accuracy of derivatized heparin and heparan sulfate (HS) glycosaminoglycan (GAG) assignments to tandem mass (MS/MS) spectra made by the first published database search application, GAG-ID. Employing a multivariate expectation-maximization algorithm, this statistical model distinguishes correct from ambiguous and incorrect database search results when computing the probability that heparin/HS GAG assignments to spectra are correct based upon database search scores...
December 9, 2016: Molecular & Cellular Proteomics: MCP
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