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https://www.readbyqxmd.com/read/28747320/microscopic-analysis-of-reconstituted-copii-coat-polymerization-and-sec16-dynamics
#1
Hirohiko Iwasaki, Tomohiro Yorimitsu, Ken Sato
COPII coat and the small GTPase Sar1 mediate protein export from the endoplasmic reticulum (ER) via specialized domains known as the ER exit sites. The peripheral ER protein Sec16 has been proposed to organize ER exit sites. However, it remains unclear how these molecules drive COPII coat polymerization. Here, we characterized the spatiotemporal relationships between the COPII components during their polymerization using fluorescence microscopy combined with an artificial planar membrane. We demonstrated that Sar1 dissociates from the membrane shortly after the COPII coat recruitment, and Sar1 is then no longer required for the COPII coat to bind to the membrane...
July 26, 2017: Journal of Cell Science
https://www.readbyqxmd.com/read/28746398/tomosyn-associates-with-secretory-vesicles-in-neurons-through-its-n-and-c-terminal-domains
#2
Cornelia J Geerts, Roberta Mancini, Ning Chen, Frank T W Koopmans, Ka Wan Li, August B Smit, Jan R T van Weering, Matthijs Verhage, Alexander J A Groffen
The secretory pathway in neurons requires efficient targeting of cargos and regulatory proteins to their release sites. Tomosyn contributes to synapse function by regulating synaptic vesicle (SV) and dense-core vesicle (DCV) secretion. While there is large support for the presynaptic accumulation of tomosyn in fixed preparations, alternative subcellular locations have been suggested. Here we studied the dynamic distribution of tomosyn-1 (Stxbp5) and tomosyn-2 (Stxbp5l) in mouse hippocampal neurons and observed a mixed diffuse and punctate localization pattern of both isoforms...
2017: PloS One
https://www.readbyqxmd.com/read/28743825/contribution-of-the-clathrin-adaptor-ap-1-subunit-%C3%A2%C2%B51-to-acidic-cluster-protein-sorting
#3
Paloma Navarro Negredo, James R Edgar, Antoni G Wrobel, Nathan R Zaccai, Robin Antrobus, David J Owen, Margaret S Robinson
Acidic clusters act as sorting signals for packaging cargo into clathrin-coated vesicles (CCVs), and also facilitate down-regulation of MHC-I by HIV-1 Nef. To find acidic cluster sorting machinery, we performed a gene-trap screen and identified the medium subunit (µ1) of the clathrin adaptor AP-1 as a top hit. In µ1 knockout cells, intracellular CCVs still form, but acidic cluster proteins are depleted, although several other CCV components were either unaffected or increased, indicating that cells can compensate for long-term loss of AP-1...
July 25, 2017: Journal of Cell Biology
https://www.readbyqxmd.com/read/28742939/disparate-peroxisome-related-defects-in-arabidopsis-pex6-and-pex26-mutants-link-peroxisomal-retrotranslocation-and-oil-body-utilization
#4
Kim L Gonzalez, Wendell A Fleming, Yun-Ting Kao, Zachary J Wright, Savina V Venkova, Meredith J Ventura, Bonnie Bartel
Catabolism of fatty acids stored in oil bodies is essential for Arabidopsis seed germination and seedling development. This fatty acid breakdown occurs in peroxisomes, organelles that sequester oxidative reactions. Import of peroxisomal enzymes is facilitated by peroxins including PEX5, a receptor that delivers cargo proteins from the cytosol to the peroxisomal matrix. After cargo delivery, a complex of the PEX1 and PEX6 ATPases and the PEX26 tail-anchored membrane protein removes ubiquitinated PEX5 from the peroxisomal membrane...
July 25, 2017: Plant Journal: for Cell and Molecular Biology
https://www.readbyqxmd.com/read/28738814/an-extended-model-of-vesicle-fusion-at-the-plasma-membrane-to-estimate-protein-lateral-diffusion-from-tirf-microscopy-images
#5
Antoine Basset, Patrick Bouthemy, Jérôme Boulanger, François Waharte, Jean Salamero, Charles Kervrann
BACKGROUND: Characterizing membrane dynamics is a key issue to understand cell exchanges with the extra-cellular medium. Total internal reflection fluorescence microscopy (TIRFM) is well suited to focus on the late steps of exocytosis at the plasma membrane. However, it is still a challenging task to quantify (lateral) diffusion and estimate local dynamics of proteins. RESULTS: A new model was introduced to represent the behavior of cargo transmembrane proteins during the vesicle fusion to the plasma membrane at the end of the exocytosis process...
July 24, 2017: BMC Bioinformatics
https://www.readbyqxmd.com/read/28736169/protein-interaction-analysis-provides-a-map-of-the-spatial-and-temporal-organization-of-the-ciliary-gating-zone
#6
Daisuke Takao, Liang Wang, Allison Boss, Kristen J Verhey
The motility and signaling functions of the primary cilium require a unique protein and lipid composition that is determined by gating mechanisms localized at the base of the cilium. Several protein complexes localize to the gating zone and may regulate ciliary protein composition; however, the mechanisms of ciliary gating and the dynamics of the gating components are largely unknown. Here, we used the BiFC (bimolecular fluorescence complementation) assay and report for the first time on the protein-protein interactions that occur between ciliary gating components and transiting cargoes during ciliary entry...
July 13, 2017: Current Biology: CB
https://www.readbyqxmd.com/read/28734834/dynein-binds-and-stimulates-axonal-motility-of-the-endosome-adaptor-and-neep21-family-member-calcyon
#7
Liang Shi, Nagendran Muthusamy, Deanna Smith, Clare Bergson
The neuron-enriched, endosomal protein Calcyon (Caly) regulates endocytosis and vesicle sorting, and is important for synaptic plasticity and brain development. In the current investigation of Caly interacting proteins in brain, the microtubule retrograde motor subunit, cytoplasmic dynein 1 heavy chain (DYNC1H), and microtubule structural proteins, α and β tubulin, were identified as Caly associated proteins by MALDI-ToF/ToF. Direct interaction of the Caly-C terminus with dynein and tubulin was further confirmed in in vitro studies...
July 19, 2017: International Journal of Biochemistry & Cell Biology
https://www.readbyqxmd.com/read/28734771/new-advances-in-autophagy-in-plants-regulation-selectivity-and-function
#8
REVIEW
Ping Wang, Yosia Mugume, Diane C Bassham
Autophagy is a major and conserved pathway for delivering unwanted proteins or damaged organelles to the vacuole for degradation and recycling. In plants, it functions as a housekeeping process to maintain cellular homeostasis under normal conditions and is induced by stress and senescence; it thus plays important roles in development, stress tolerance and metabolism. Autophagy can both execute bulk degradation and be highly selective in targeting cargos under specific environmental conditions or during certain developmental processes...
July 19, 2017: Seminars in Cell & Developmental Biology
https://www.readbyqxmd.com/read/28730938/stratum-corneum-modulation-by-chemical-enhancers-and-lipid-nanostructures-implications-for-transdermal-drug-delivery
#9
Mudra Saurabh Kapoor, Shruti GuhaSarkar, Rinti Banerjee
Skin is the outermost and largest protective covering of the body. The uppermost layer of the skin, stratum corneum also called the horny layer is composed of keratin-filled cells covered by a lipid matrix which shields the skin from physical and chemical entrants. The lipid lamellar structure comprises of ceramides, cholesterol, fatty acids and proteins. Chemical enhancers that mimic the lamellar chemistry, reversibly fluidize the latter can be utilized for enhancing transport of cargo across the epidermis into the dermis...
July 2017: Therapeutic Delivery
https://www.readbyqxmd.com/read/28727280/two-novel-effectors-of-trafficking-and-maturation-of-the-yeast-plasma-membrane-h-atpase
#10
Yosef Geva, Jonathan Crissman, Eric C Arakel, Natalia Gómez-Navarro, Silvia G Chuartzman, Kyle R Stahmer, Blanche Schwappach, Elizabeth A Miller, Maya Schuldiner
The endoplasmic reticulum (ER) is the entry site of proteins into the endomembrane system. Proteins exit the ER via COPII vesicles in a selective manner, mediated either by direct interaction with the COPII coat or aided by cargo receptors. Despite the fundamental role of such receptors in protein sorting, only a few have been identified. To further define the machinery that packages secretory cargo and targets proteins from the ER to Golgi membranes, we used multiple systematic approaches, which revealed two uncharacterized proteins that mediate the trafficking and maturation of Pma1, the essential yeast plasma membrane proton ATPase...
July 20, 2017: Traffic
https://www.readbyqxmd.com/read/28725673/efficiency-data-of-intracellular-recombinant-protein-delivery-using-cationic-lipid-coated-silk-fibroin-particle
#11
Woo-Jin Kim, Bong-Soo Kim, Young-Dan Cho, Won-Joon Yoon, Jeong-Hwa Baek, Kyung-Mi Woo, Hyun-Mo Ryoo
This article presents data related to the research article "Fibroin particle-supported cationic lipid layers for highly efficient intracellular protein delivery" and focuses on the delivery efficiency aspects of the fibroin particle-cationic lipid complex (Fibroplex), including its fabrication and the intracellular delivery to the mouse skin tissue. We introduced a stable lipid-particle complex called "Fibroplex", formed by loading cargo protein onto a silk fibroin spherical particle core complexed with cationic liposomes to address the intracellular recombinant protein delivery...
August 2017: Data in Brief
https://www.readbyqxmd.com/read/28725021/bovine-milk-derived-exosomes-from-colostrum-are-enriched-with-proteins-implicated-in-immune-response-and-growth
#12
Monisha Samuel, David Chisanga, Michael Liem, Shivakumar Keerthikumar, Sushma Anand, Ching-Seng Ang, Christopher G Adda, Ellen Versteegen, Markandeya Jois, Suresh Mathivanan
Exosomes are extracellular vesicles secreted by multiple cell types into the extracellular space. They contain cell-state specific cargos which often reflects the (patho)physiological condition of the cells/organism. Milk contains high amounts of exosomes and it is unclear whether their cargo is altered based on the lactation stage of the organism. Here, we isolated exosomes from bovine milk that were obtained at various stages of lactation and examined the content by quantitative proteomics. Exosomes were isolated by OptiPrep density gradient centrifugation from milk obtained from cow after 24, 48 and 72 h post calving...
July 19, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28724787/disrupted-apical-exocytosis-of-cargo-vesicles-causes-enteropathy-in-fhl5-patients-with-munc18-2-mutations
#13
Georg F Vogel, Jorik M van Rijn, Iris M Krainer, Andreas R Janecke, Carsten Posovzsky, Marta Cohen, Claire Searle, Prevost Jantchou, Johanna C Escher, Natalie Patey, Ernest Cutz, Thomas Müller, Sabine Middendorp, Michael W Hess, Lukas A Huber
Familial hemophagocytic lymphohistiocytosis 5 (FHL5) is an autosomal recessive disease caused by mutations in STXBP2, coding for Munc18-2, which is required for SNARE-mediated membrane fusion. FHL5 causes hematologic and gastrointestinal symptoms characterized by chronic enteropathy that is reminiscent of microvillus inclusion disease (MVID). However, the molecular pathophysiology of FHL5-associated diarrhea is poorly understood. Five FHL5 patients, including four previously unreported patients, were studied...
July 20, 2017: JCI Insight
https://www.readbyqxmd.com/read/28724525/trim37-a-novel-e3-ligase-for-pex5-mediated-peroxisomal-matrix-protein-import
#14
Wei Wang, Zhi-Jie Xia, Jean-Claude Farré, Suresh Subramani
Most proteins destined for the peroxisomal matrix depend on the peroxisomal targeting signals (PTSs), which require the PTS receptor PEX5, whose deficiency causes fatal human peroxisomal biogenesis disorders (PBDs). TRIM37 gene mutations cause muscle-liver-brain-eye (mulibrey) nanism. We found that TRIM37 localizes in peroxisomal membranes and ubiquitylates PEX5 at K464 by interacting with its C-terminal 51 amino acids (CT51), which is required for PTS protein import. PEX5 mutations (K464A or ΔCT51), or TRIM37 depletion or mutation, reduce PEX5 abundance by promoting its proteasomal degradation, thereby impairing its functions in cargo binding and PTS protein import in human cells...
July 19, 2017: Journal of Cell Biology
https://www.readbyqxmd.com/read/28723420/plant-and-yeast-cornichon-possess-a-conserved-acidic-motif-required-for-correct-targeting-of-plasma-membrane-cargos
#15
Paul Rosas-Santiago, Daniel Lagunas-Gomez, Carolina Yáñez-Domínguez, Rosario Vera-Estrella, Olga Zimmermannova, Hana Sychrová, Omar Pantoja
The export of membrane proteins along the secretory pathway is initiated at the endoplasmic reticulum after proteins are folded and packaged inside this organelle by their recruiting into the coat complex COPII vesicles. It is proposed that cargo receptors are required for the correct transport of proteins to its target membrane, however, little is known about ER export signals for cargo receptors. Erv14/Cornichon belong to a well conserved protein family in Eukaryotes, and have been proposed to function as cargo receptors for many transmembrane proteins...
July 16, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28722341/proteomic-analysis-of-neuroblastoma-derived-exosomes-new-insights-into-a-metastatic-signature
#16
Marta Colletti, Andrea Petretto, Angela Galardi, Virginia Di Paolo, Luigi Tomao, Chiara Lavarello, Elvira Inglese, Maurizio Bruschi, Ana Amor Lopez, Luisa Pascucci, Birgit Geoerger, Hector Peinado, Locatelli Franco, Di Giannatale Angela
Neuroblastoma (NB) is the most common extracranial pediatric solid tumor. Around 70% of patients with metastatic disease at diagnosis present bone-marrow infiltration, which is considered a marker of poor outcome; however, the mechanism underlying this specific tropism has to be elucidated. Tumor-derived exosomes may support metastatic progression in several tumors by interacting with the microenvironment, and may serve as tumor biomarkers. The main objective of this study was to identify an exosomal signature associated with NB metastatic bone-marrow dissemination...
July 19, 2017: Proteomics
https://www.readbyqxmd.com/read/28722016/silk-micrococoons-for-protein-stabilisation-and-molecular-encapsulation
#17
Ulyana Shimanovich, Francesco S Ruggeri, Erwin De Genst, Jozef Adamcik, Teresa P Barros, David Porter, Thomas Müller, Raffaele Mezzenga, Christopher M Dobson, Fritz Vollrath, Chris Holland, Tuomas P J Knowles
Naturally spun silks generate fibres with unique properties, including strength, elasticity and biocompatibility. Here we describe a microfluidics-based strategy to spin liquid native silk, obtained directly from the silk gland of Bombyx mori silkworms, into micron-scale capsules with controllable geometry and variable levels of intermolecular β-sheet content in their protein shells. We demonstrate that such micrococoons can store internally the otherwise highly unstable liquid native silk for several months and without apparent effect on its functionality...
July 19, 2017: Nature Communications
https://www.readbyqxmd.com/read/28720840/positioning-of-the-scrambled-receptor-requires-udp-glc-sterol-glucosyltransferase-80b1-in-arabidopsis-roots
#18
Victoria G Pook, Meera Nair, KookHui Ryu, James C Arpin, John Schiefelbein, Kathrin Schrick, Seth DeBolt
The biological function of sterol glucosides (SGs), the most abundant sterol derivatives in higher plants, remains uncertain. In an effort to improve our understanding of these membrane lipids we examined phenotypes exhibited by the roots of Arabidopsis (Arabidopsis thaliana) lines carrying insertions in the UDP-Glc:sterol glucosyltransferase genes, UGT80A2 and UGT80B1. We show that although ugt80A2 mutants exhibit significantly lower levels of total SGs they are morphologically indistinguishable from wild-type plants...
July 18, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28720664/dynein-efficiently-navigates-the-dendritic-cytoskeleton-to-drive-the-retrograde-trafficking-of-bdnf-trkb-signaling-endosomes
#19
Swathi Ayloo, Pedro Guedes-Dias, Amy E Ghiretti, Erika L F Holzbaur
The efficient transport of cargoes within axons and dendrites is critical for neuronal function. While we have a basic understanding of axonal transport, much less is known about transport in dendrites. We employed an optogenetic approach to recruit motor proteins to cargo in real time within axons or dendrites in hippocampal neurons. Kinesin-1, a robust axonal motor, moves cargo less efficiently in dendrites. In contrast, cytoplasmic dynein efficiently navigates both axons and dendrites; in both compartments, dynamic microtubule plus-ends enhance dynein-dependent transport...
July 18, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28718761/the-human-cytoplasmic-dynein-interactome-reveals-novel-activators-of-motility
#20
Willam B Redwine, Morgan E DeSantis, Ian Hollyer, Zaw Min Htet, Phuoc Tien Tran, Selene K Swanson, Laurence Florens, Michael P Washburn, Samara L Reck-Peterson
In human cells, cytoplasmic dynein-1 is essential for long-distance transport of many cargos, including organelles, RNAs, proteins, and viruses, towards microtubule minus ends. To understand how a single motor achieves cargo specificity, we identified the human dynein interactome by attaching a promiscuous biotin ligase ("BioID") to seven components of the dynein machinery, including a subunit of the essential cofactor dynactin. This method reported spatial information about the large cytosolic dynein/dynactin complex in living cells...
July 18, 2017: ELife
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