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Noah Fine, Ioannis D Dimitriou, Jacob Rullo, María José Sandí, Björn Petri, Jack Haitsma, Hisham Ibrahim, Jose La Rose, Michael Glogauer, Paul Kubes, Myron Cybulsky, Robert Rottapel
Leukocyte crawling and transendothelial migration (TEM) are potentiated by shear stress caused by blood flow. The mechanism that couples shear stress to migration has not been fully elucidated. We found that mice lacking GEF-H1 (GEF-H1(-/-)), a RhoA-specific guanine nucleotide exchange factor (GEF), displayed limited migration and recruitment of neutrophils into inflamed tissues. GEF-H1(-/-) leukocytes were deficient in in vivo crawling and TEM in the postcapillary venules. We demonstrated that although GEF-H1 deficiency had little impact on the migratory properties of neutrophils under static conditions, shear stress triggered GEF-H1-dependent spreading and crawling of neutrophils and relocalization of GEF-H1 to flotillin-2-rich uropods...
October 10, 2016: Journal of Cell Biology
Timothy Wai, Shotaro Saita, Hendrik Nolte, Sebastian Müller, Tim König, Ricarda Richter-Dennerlein, Hans-Georg Sprenger, Joaquin Madrenas, Mareike Mühlmeister, Ulrich Brandt, Marcus Krüger, Thomas Langer
The SPFH (stomatin, prohibitin, flotillin, HflC/K) superfamily is composed of scaffold proteins that form ring-like structures and locally specify the protein-lipid composition in a variety of cellular membranes. Stomatin-like protein 2 (SLP2) is a member of this superfamily that localizes to the mitochondrial inner membrane (IM) where it acts as a membrane organizer. Here, we report that SLP2 anchors a large protease complex composed of the rhomboid protease PARL and the i-AAA protease YME1L, which we term the SPY complex (for SLP2-PARL-YME1L)...
October 13, 2016: EMBO Reports
Gerald Grandl, Sebastian Müller, Hansjörg Moest, Caroline Moser, Bernd Wollscheid, Christian Wolfrum
OBJECTIVE: Adipose tissue shows a high degree of plasticity, and adipocyte hyperplasia is an important mechanism for adipose tissue expansion. Different adipose depots respond differently to an increased demand for lipid storage. Orchestrating cellular expansion in vivo requires extracellular matrix (ECM) remodeling and a high degree of interaction between cells and ECM. METHODS: We studied decellularized primary adipose stromal cell derived ECM of different adipose depots and reseeded them with primary adipose precursors...
October 2016: Molecular Metabolism
Yuqian Luo, Takeshi Akama, Akiko Okayama, Aya Yoshihara, Mariko Sue, Kenzaburo Oda, Moyuru Hayashi, Yuko Ishido, Hisashi Hirano, Naoki Hiroi, Ryohei Katoh, Koichi Suzuki
BACKGROUND: Thyroglobulin (Tg) stored in thyroid follicles regulates follicular function in thyroid hormone (TH) synthesis by suppressing thyroid-specific gene expression in a concentration-dependent manner. Thus, Tg is an intrinsic negative-feedback regulator that can restrain the effect of thyroid stimulating hormone (TSH) in the follicle. However, the underlying mechanisms by which Tg exerts its prominent autoregulatory effect following recognition by thyrocytes remains unclear. METHODS: To identify potential proteins that recognize and interact with Tg, mass spectrometry (MS) was used to analyze immunoprecipitated Tg-bound proteins derived from Tg-treated rat thyroid FRTL-5 cells...
September 27, 2016: Thyroid: Official Journal of the American Thyroid Association
Ling Wang, Tore Skotland, Viktor Berge, Kirsten Sandvig, Alicia Llorente
Exosomes have recently appeared as a novel source of non-invasive cancer biomarkers since tumor-specific molecules can be found in exosomes isolated from biological fluids. We have previously analyzed the proteome of urinary exosomes by mass spectrometry, and identified proteins differentially expressed in prostate cancer patients compared to healthy males. Since mass spectrometry is so far not commonly used in clinical laboratories, we have here investigated whether antibody-based methods such as Western blot or ELISA can be used to validate the use of the identified proteins as prostate cancer biomarkers...
September 21, 2016: European Journal of Pharmaceutical Sciences
Martijn J C van Herwijnen, Marijke I Zonneveld, Soenita Goerdayal, Esther N M Nolte-'t Hoen, Johan Garssen, Bernd Stahl, A F Maarten Altelaar, Frank A Redegeld, Marca H M Wauben
Breast milk contains several macromolecular components with distinctive functions, whereby milk fat globules and casein micelles mainly provide nutrition to the newborn, while whey contains molecules that can stimulate the newborn's developing immune system and gastrointestinal tract. Although extracellular vesicles (EV) have been identified in breast milk, their physiological function and composition has not been addressed in detail. EV are submicron sized vehicles released by cells for intercellular communication via selectively incorporated lipids, nucleic acids and proteins...
September 6, 2016: Molecular & Cellular Proteomics: MCP
Yuan Liu, Xiaodong Guo, Liyuan Wu, Mei Yang, Zhiwei Li, Yinjie Gao, Shuhong Liu, Guangde Zhou, Jingmin Zhao
: Hepatocellular carcinoma (HCC) occurs predominantly in patients with underlying chronic liver disease and cirrhosis. Toll-like receptors (TLRs) play an important role in innate immune responses and TLR signaling has been associated with various chronic liver diseases. Lipid rafts provide the necessary microenvironment for certain specialized signaling events to take place, such as the innate immune recognition. The purpose of this study was to determine the pattern of TLR7 expression in HCC, how to recruit TLR7 into lipid rafts responded to ligands and whether targeting TLR7 might have beneficial effects...
August 30, 2016: Oncotarget
Mihael Lazebnik, Rahul K Keswani, Daniel W Pack
PURPOSE: siRNA may be delivered as electrostatic complexes with cationic lipids (lipoplexes) or polycations (polyplexes). The purpose of this project was to determine the effect of cellular internalization mechanism(s) on siRNA-mediated gene silencing efficiency. METHODS: Lipoplexes were formed comprising siRNA and N-[1-(2,3-dioleoyloxy)propyl]-N,N,N-trimethylammonium methyl-sulfate (DOTAP), cholesterol and dioleoyl phosphatidylethanolamine (DOPE), and polyplexes comprised siRNA with polyethylenimine (PEI)...
September 1, 2016: Pharmaceutical Research
Nadine Legros, Stefanie Dusny, Hans-Ulrich Humpf, Gottfried Pohlentz, Helge Karch, Johannes Müthing
Shiga toxin (Stx)-mediated injury to microvascular endothelial cells in the brain significantly contributes to the pathogenesis of the hemolytic-uremic syndrome caused by enterohemorrhagic Escherichia coli (EHEC). Stxs are AB5 toxins and the B-pentamers of the two major Stx subtypes Stx1a and Stx2a preferentially bind to the glycosphingolipid (GSL) globotriaosylceramide (Gb3Cer) expressed by human endothelial cells. Here we report on comprehensive structural analysis of the different lipoforms of Gb3Cer (Galα4Galβ4Glcβ1Cer) and globotetraosylceramide (Gb4Cer, GalNAcβ3Galα4Galβ4Glcβ1Cer, the less effective Stx receptor) of primary human brain microvascular endothelial cells and their association with lipid rafts...
August 24, 2016: Glycobiology
Qin Yang, Min Zhu, Zi Wang, Hui Li, Weihua Zhou, Xiaojuan Xiao, Bin Zhang, Weixin Hu, Jing Liu
The membrane-cytoskeletal protein 4.1N has recently been proposed as a tumor suppressor in a number of cancers of epithelial origin, including non-small-cell lung cancer (NSCLC). However, the molecular mechanism associated with 4.1N tumor suppression remains has not been thoroughly characterized. In this study, 4.1N was shown to directly interact with the lipid raft marker flotillin-1 through its FERM and U2 domains in several different NSCLC cell lines using immunoprecipitation, co-immunoprecipitation and pull-down assays...
July 22, 2016: Tumour Biology: the Journal of the International Society for Oncodevelopmental Biology and Medicine
Mohammad Abdullah, Hiroshi Takase, Mari Nunome, Hiroyuki Enomoto, Jin-Ichi Ito, Jian-Sheng Gong, Makoto Michikawa
Exosomes are small extracellular vesicles secreted by variety of cell types such as neurons, astrocytes, and oligodendrocytes. It is suggested that exosomes play essential role in the maintenance of the neuronal functions and also in the clearance of amyloid-β (Aβ) from the brain. Aβ is well known to cause neuronal cell death, whereas little is known about its effect on astrocytes. In this study, we examined the effect of Aβ on release of exosomes from astrocytes in culture. We analyzed release of exosomes and apoE, both of which are known to remove/clear Aβ from the brain, in the culture medium of astrocytes...
July 2, 2016: Journal of Alzheimer's Disease: JAD
Felix Dempwolff, Felix K Schmidt, Ana B Hervás, Alex Stroh, Thomas C Rösch, Cornelius N Riese, Simon Dersch, Thomas Heimerl, Daniella Lucena, Nikola Hülsbusch, Claudia A O Stuermer, Norio Takeshita, Reinhard Fischer, Bruno Eckhardt, Peter L Graumann
Biological membranes have been proposed to contain microdomains of a specific lipid composition, in which distinct groups of proteins are clustered. Flotillin-like proteins are conserved between pro-and eukaryotes, play an important function in several eukaryotic and bacterial cells, and define in vertebrates a type of so-called detergent-resistant microdomains. Using STED microscopy, we show that two bacterial flotillins, FloA and FloT, form defined assemblies with an average diameter of 85 to 110 nm in the model bacterium Bacillus subtilis...
June 2016: PLoS Genetics
Jin-Mo Kim, Seon-Heui Cha, Yu Ree Choi, Ilo Jou, Eun-Hye Joe, Sang Myun Park
Parkinson's disease (PD) is a common chronic and progressive neurodegenerative disorder. Although the cause of PD is still poorly understood, mutations in many genes including SNCA, parkin, PINK1, LRRK2, and DJ-1 have been identified in the familial forms of PD. It was recently proposed that alterations in lipid rafts may cause the neurodegeneration shown in PD. Here, we observe that DJ-1 deficiency decreased the expression of flotillin-1 (flot-1) and caveolin-1 (cav-1), the main protein components of lipid rafts, in primary astrocytes and MEF cells...
2016: Scientific Reports
Frauke Völlner, Jawahir Ali, Nina Kurrle, Yvonne Exner, Rüdiger Eming, Michael Hertl, Antje Banning, Ritva Tikkanen
Desmosomes are adhesion plaques that mediate cell-cell adhesion in many tissues, including the epidermis, and generate mechanical resistance to tissues. The extracellular domains of desmosomal cadherin proteins, desmogleins and desmocollins, are required for the interaction with cadherins of the neighbouring cells, whereas their cytoplasmic tails associate with cytoplasmic proteins which mediate connection to intermediate filaments. Disruption of desmosomal adhesion by mutations, autoantibodies or bacterial toxins results in severe human disorders of e...
2016: Scientific Reports
Luciane Martins, Adriana Franco Paes Leme, Kamila Rosamilia Kantovitz, Enilson Antonio Sallum, Márcio Zaffalon Casati, Francisco Humberto Nociti Junior
Basic, pre-clinical and clinical studies have documented the potential of amelogenin, and its variants, to affect cell response and tissue regeneration. However, the mechanisms are unclear. Thus, the aim of the present study was to identify, in cementoblasts, novel binding partners for an alternatively spliced amelogenin form (Leucine-Rich Amelogenin Peptide - LRAP), which is supposed to act as a signaling molecule in epithelial-mesenchymal interactions. LRAP-binding protein complexes from immortalized murine cementoblasts (OCCM-30) were achieved by capture affinity assay (GST pull down) and proteins present in these complexes were identified by mass spectrometry and immunoblotting...
June 9, 2016: Journal of Cellular Physiology
Xuri Mou, Shuliang Liu
Lung adenocarcinoma, as a common form of non-small cell lung cancer, poses a significant threat to public health worldwide. Previous studies have reported that flotillin-2 (Flot2) is often overexpressed in various tumors and is h correlated with tumor progression and patient survival. Dysregulated microRNA (miRNA) is associated with various cancers, including lung adenocarcinoma. However, little is known about the miRNAs targeting Flot2 in lung adenocarcinoma. In this study, we found that the expression level of miR-485 was downregulated in four lung adenocarcinoma cell lines and tissues and that the reduced miR-485 expression was associated with tumor metastasis...
September 2, 2016: Biochemical and Biophysical Research Communications
Moon-Yong Park, Nahyun Kim, Li-Ling Wu, Guang-Yan Yu, Kyungpyo Park
Endocytosis has numerous functions in cellular homeostasis. Defects in the endocytic pathway of receptors may lead to dysfunction of salivary gland secretion. Therefore, elucidating the complex mechanisms of endocytosis may facilitate solutions for disease treatment and prevention. The muscarinic type 3 receptor (M3R), a G-protein-coupled receptor (GPCR) located in the plasma membrane, is involved in numerous physiological activities such as smooth muscle contraction and saliva secretion. M3R enters cells through clathrin-mediated endocytosis (CME), while flotillins (flot-1 and -2), highly conserved proteins residing in lipid-raft microdomains, make use of clathrin-independent endocytosis (CIE) for their internalization...
August 5, 2016: Biochemical and Biophysical Research Communications
Ya Niu, Ziyun Shao, Hui Wang, Jiaqi Yang, Feifei Zhang, Yuhao Luo, Lijun Xu, Yanqing Ding, Liang Zhao
LIM and SH3 protein 1(LASP1) can promote colorectal cancer (CRC) progression and metastasis, but the mechanism remains unclear. Here, we show that LASP1 interacts with S100 calcium binding protein A11(S100A11) and enhances its expression in CRC. LASP1-S100A11 axis is essential for TGFβ-mediated epithelial-mesenchymal transition (EMT) and cell aggressive phenotype. Clinically, S100A11 is overexpressed in CRC tissues and localized in both the cytoplasm and the nucleus of CRC cells. Overexpression of S100A11 in cytoplasmic and nuclear subcellular compartments is associated with tumor metastasis and poor prognosis of CRC patients...
2016: Scientific Reports
Shingo Enomoto, Kunio Shimizu, Masashi Nibuya, Hiroyuki Toda, Aihide Yoshino, Eiji Suzuki, Takashi Kondo, Hiroshi Fukuda
Although electroconvulsive therapy (ECT) is clinically used for severe depression and drug-resistant Parkinson's disease, its exact biological background and mechanism have not yet been fully elucidated. Two potential explanations have been presented so far to explain the increased neuroplastic and resilient profiles of multiple ECT administrations. One is the alteration of central neurotransmitter receptor densities and the other is the expressional upregulation of brain derived neurotrophic factor in various brain regions with enhanced hippocampal neurogenesis and mossy fiber sprouting...
June 15, 2016: Neuroscience Letters
Yen-Ning Huang, Ching-I Lin, Hsiang Liao, Chin-Yu Liu, Yue-Hua Chen, Wan-Chun Chiu, Shyh-Hsiang Lin
Epidemiological investigations have shown that Alzheimer's disease (AD) is one of the most common neurodegenerative diseases. It has been indicated that the cholesterol concentration in the brain of AD patients is higher than that in normal people. In this study, we investigated the effects of cholesterol concentrations, 0, as the control, 3.125, 12.5, and 25μM, on cholesterol metabolism, neuron survival, AD-related protein expressions, and cell morphology and apoptosis using SH-SY5Y human neuroblastoma cells...
July 22, 2016: Neuroscience
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