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clathrin endocytosis

Shengwei Jiang, Yuchun Lin, Huan Yao, Chuanli Yang, Liyin Zhang, Bing Luo, Zhao Lei, Liwei Cao, Naibo Lin, Xiangyang Liu, Zhongning Lin, Chengyong He
Unfolded protein response (UPR) and endoplasmic reticulum (ER)-phagy are essential for cell homeostasis. Quantum dots (QDs), which have been widely used for biomedical applications, can accumulate in the kidney tissues and may cause renal dysfunction. However, the molecular mechanism of QDs-induced nephrotoxicity is still obscure. The present study was aimed to elucidate the role and mechanism of UPR and ER-phagy in QDs-induced nephrotoxicity. Herein, human embyronic kidney (HEK) cells were exposed to 15, 30, 45, and 60 nM cadmium telluride (CdTe)-QDs for 12 and 24 h...
February 12, 2018: Archives of Toxicology
N Beztsinna, M B C de Matos, J Walther, C Heyder, E Hildebrandt, G Leneweit, E Mastrobattista, R J Kok
Ribosome inactivating proteins (RIPs) are highly potent cytotoxins that have potential as anticancer therapeutics. Mistletoe lectin 1 (ML1) is a heterodimeric cytotoxic protein isolated from European Mistletoe and belongs to RIP class II. The aim of this project was to systematically study ML1 cell binding, endocytosis pathway(s), subcellular processing and apoptosis activation. For this purpose, state of the art cell imaging equipment and automated image analysis algorithms were used. ML1 displayed very fast binding to sugar residues on the membrane and energy-dependent uptake in CT26 cells...
February 9, 2018: Scientific Reports
Yifei Guo, Shuang Zhao, Hanhong Qiu, Ting Wang, Yanna Zhao, Meihua Han, Zhengqi Dong, Xiangtao Wang
It was reported that the shape of nanocarriers played an important role in achieving better therapeutic effect. To optimize the morphology and enhance the antitumor efficacy, in this study based on the amphiphilic PAMAM-b-OEG codendrimer (POD), docetaxel-loaded spherical and flake-like nanoparticles (DTX nanospheres and nanosheets) were prepared via antisolvent precipitation method with the similar particle size, surface charge, stability, and release profiles. The feed weight ratio of DTX/POD and the branched structure of OEG dendron were suggested to influence on the shapes of the self-assembled nanostructures...
February 9, 2018: Bioconjugate Chemistry
Mónica Loreto Acevedo, Francisco García-de Gracia, Camila Miranda-Cárdenas, Ricardo Soto-Rifo, Francisco Aguayo, Oscar León
Self-inactivating VSVG-pseudotyped murine leukemia virus (SIN-VSVG-MLV) has been widely used to generate stable cell lines and produce gene delivery vectors. Despite the broad cellular tropism of the VSVG-pseudotyped MLV, we observed differential viral transduction efficiency depending on the host cell type used. In order to determine the mechanism underlying these differences, we used a GFP-expressing SIN-VSVG-MLV and analyzed the major steps of viral transduction in different cell lines including human epithelial, T-lymphocytes, monocytes and murine fibroblast cells...
February 6, 2018: Journal of Virological Methods
Laetitia Bertot, Alexandre Grassart, Thibault Lagache, Giacomo Nardi, Cyril Basquin, Jean-Christophe Olivo-Marin, Nathalie Sauvonnet
Eukaryotic cells internalize cargos specifically through clathrin-mediated endocytosis (CME) or clathrin-independent endocytosis (CIE). EndophilinA2 was shown as preferentially implicated in CIE, although initially involved in CME. Here, we investigated the native interplay of endophilinA2 and dynamin2 during CME as compared to CIE. We developed an unbiased integrative approach based on genome engineering, robust tracking methodology, and advanced analytics. We statistically identified CME and CIE subpopulations corresponding to abortive, active, and static endocytic events...
February 6, 2018: Cell Reports
Wen Song, Chuanxu Yang, Dang Quang Svend Le, Yumei Zhang, Jørgen Kjems
Controlling mesenchymal stem cells (MSCs) differentiation by RNA interference (RNAi) is a promising approach for next-generation regenerative medicine. However, efficient delivery of RNAi therapeutics is still a limiting factor. In this study, we have developed a simple, biocompatible and highly effective delivery method of small RNA therapeutics into hMSCs from an implant surface by calcium ions. First, we demonstrated that simple Ca/siGFP nanocomplexes were able to efficiently silence GFP in GFP-expressing hMSCs with adequate Ca2+ concentration (>5 mM)...
February 8, 2018: ACS Applied Materials & Interfaces
Marko Kaksonen, Aurélien Roux
Clathrin-mediated endocytosis is a key process in vesicular trafficking that transports a wide range of cargo molecules from the cell surface to the interior. Clathrin-mediated endocytosis was first described over 5 decades ago. Since its discovery, over 50 proteins have been shown to be part of the molecular machinery that generates the clathrin-coated endocytic vesicles. These proteins and the different steps of the endocytic process that they mediate have been studied in detail. However, we still lack a good understanding of how all these different components work together in a highly coordinated manner to drive vesicle formation...
February 7, 2018: Nature Reviews. Molecular Cell Biology
Eun-Jeong Choi, Hyun Gee Lee, Il-Hong Bae, Wanil Kim, Jungwon Park, Tae Ryong Lee, Eun-Gyung Cho
Acne vulgaris is an inflammatory disease occurring in the pilosebaceous unit and is the most common skin condition in young people. A gram-positive bacterium, Propionibacterium acnes (P. acnes), has been suspected to contribute to the development of acne. Here, we report that P. acnes constitutively releases extracellular vesicles (EVs) exhibiting typical EV morphology and size. Moreover, the P. acnes-derived EVs (PEVs) can induce acne-like phenotypes in human epidermal keratinocytes and a reconstituted human skin model...
January 30, 2018: Journal of Investigative Dermatology
Li-Li Huang, Li-Li Wu, Xue Li, Kejiang Liu, Dongxu Zhao, Hai-Yan Xie
Entry is the first and critical step of viral infection, while the entry mechanisms of many viruses are still unclear due to the lack of efficient technology. In this report, by taking advantage of the single-virion fluorescence tracking technique and simultaneous dual-labeling methods for viruses, we developed; the entry pathway of vaccinia virus from tiantan strain (VACV-TT) was studied in real-time. By combining with the technologies of virology and cell biology, we found that VACV-TT moved toward the Vero cell body along the filopodia induced by the virions interaction, and then, they were internalized through macropinocytosis, which was an actin-, ATP-dependent but clathrin-, caveolin-independent endocytosis...
February 2, 2018: Analytical Chemistry
Xing Chen, Ziheng Liu, Zelin Shan, Weiyi Yao, Aihong Gu, Wenyu Wen
Traffic of cargo across membranes helps establish, maintain, and reorganize distinct cellular compartments and is fundamental to many metabolic processes. The cargo-selective endocytic adaptor Numb participates in clathrin-dependent endocytosis by attaching cargoes to the clathrin adaptor α-adaptin. The phosphorylation of Numb at Ser-265 and Ser-284 recruits the regulatory protein 14-3-3, accompanied by the dissociation of Numb from α-adaptin and Numb's translocation from the cortical membrane to the cytosol...
January 30, 2018: Journal of Biological Chemistry
Brandon L Scott, Kem A Sochacki, Shalini T Low-Nam, Elizabeth M Bailey, QuocAhn Luu, Amy Hor, Andrea M Dickey, Steve Smith, Jason G Kerkvliet, Justin W Taraska, Adam D Hoppe
Clathrin-mediated endocytosis (CME) internalizes plasma membrane by reshaping small regions of the cell surface into spherical vesicles. The key mechanistic question of how coat assembly produces membrane curvature has been studied with molecular and cellular structural biology approaches, without direct visualization of the process in living cells; resulting in two competing models for membrane bending. Here we use polarized total internal reflection fluorescence microscopy (pol-TIRF) combined with electron, atomic force, and super-resolution optical microscopy to measure membrane curvature during CME...
January 29, 2018: Nature Communications
Nauman Rahim Khan, Tin Wui Wong
This study focuses on the use of ethosome and microwave technologies to facilitate skin penetration and/or deposition of 5-fluorouracil in vitro and in vivo. Low ethanol ethosomes were designed and processed by mechanical dispersion technique and had their size, zeta potential, morphology, drug content and encapsulation efficiency characterized. The skin was pre-treated with microwave at 2450 MHz for 2.5 min with ethosomes applied topically and subjected to in vitro and in vivo skin drug permeation as well as retention evaluation...
January 29, 2018: Artificial Cells, Nanomedicine, and Biotechnology
Changling Sun, Xiaoying Li, Xiaodong Du, Teng Wang
In this study, we reported a redox-responsive drug delivery system (DDS) based on heparosan and deoxycholic acid conjugates (HSDs) for effective treatment of laryngopharyngeal carcinoma. The amphiphilic HSDs can self-assemble into stable nanoscale micelles in aqueous medium with favorable drug loading capacity for doxorubicin (DOX). The HSD micelles can exhibit glutathione (GSH)-triggered drug release behavior and reach a nearly 100% release rate in a high GSH level (10 mM) environment. Moreover, FaDu cancer cells can internalize HSD micelles by clathrin-mediated endocytosis, which is energy dependent, fast, and effective...
January 22, 2018: International Journal of Biological Macromolecules
Wen-Qiang Xia, Yan Liang, Yao Chi, Li-Long Pan, Jing Zhao, Shu-Sheng Liu, Xiao-Wei Wang
Begomoviruses are exclusively transmitted by whiteflies in a persistent circulative manner and cause considerable economic losses to crop production worldwide. Previous studies have shown that begomoviruses accumulate in vesicle-like structures in whitefly midgut cells and that clathrin-mediated endocytosis is responsible for their internalization. However, the process by which begomoviruses are trafficked within whitefly midgut cells remains largely unknown. In this study, we investigated the roles of vesicle trafficking in the transport of Tomato yellow leaf curl virus (TYLCV), a begomovirus that has spread to over 50 countries and caused extensive damage to a range of important crops, within midgut cells of whitefly (Bemisia tabaci)...
January 25, 2018: PLoS Pathogens
Syed A Morshed, Risheng Ma, Rauf Latif, Terry F Davies
The thyroid-stimulating hormone receptor (TSHR) is a heterotrimeric guanine nucleotide-binding protein (G protein)-coupled receptor (GPCR). Autoimmune hyperthyroidism, commonly known as Graves' disease (GD), is caused by stimulating autoantibodies to the TSHR. We previously described TSHR-specific antibodies (TSHR-Abs) in GD that recognize linear epitopes in the cleavage region of the TSHR ectodomain (C-TSHR-Abs) and induce thyroid cell apoptosis instead of stimulating the TSHR. We found that C-TSHR-Abs entered the cell through clathrin-mediated endocytosis but did not trigger endosomal maturation and failed to undergo normal vesicular sorting and trafficking...
January 23, 2018: Science Signaling
Maria M Garcia-Alai, Johannes Heidemann, Michal Skruzny, Anna Gieras, Haydyn D T Mertens, Dmitri I Svergun, Marko Kaksonen, Charlotte Uetrecht, Rob Meijers
In clathrin-mediated endocytosis, adapter proteins assemble together with clathrin through interactions with specific lipids on the plasma membrane. However, the precise mechanism of adapter protein assembly at the cell membrane is still unknown. Here, we show that the membrane-proximal domains ENTH of epsin and ANTH of Sla2 form complexes through phosphatidylinositol 4,5-bisphosphate (PIP2) lipid interfaces. Native mass spectrometry reveals how ENTH and ANTH domains form assemblies by sharing PIP2 molecules...
January 23, 2018: Nature Communications
Anuttoma Ray, Parul Katoch, Nimansha Jain, Parmender P Mehta
Defects in assembly of gap junction forming proteins, called connexin (Cx)s, are observed in a variety of cancers. Connexin32 is expressed by the polarized cells in epithelia. We discovered two dileucine-based motifs, which govern the intracellular sorting and endocytosis of transmembrane proteins, in the carboxyl tail of Cx32 and explored their role in regulating its endocytosis and gap junction forming ability in pancreatic and prostate cancer cells. One motif, designated as LI, was located near the juxtamembrane domain whereas the other, designated as LL, distally...
January 19, 2018: Journal of Cell Science
Tetsuya Takeda, Toshiya Kozai, Huiran Yang, Daiki Ishikuro, Kaho Seyama, Yusuke Kumagai, Tadashi Abe, Hiroshi Yamada, Takayuki Uchihashi, Toshio Ando, Kohji Takei
Dynamin is a mechanochemical GTPase essential for membrane fission during clathrin-mediated endocytosis. Dynamin forms helical complexes at the neck of clathrin-coated pits and their structural changes coupled with GTP hydrolysis drive membrane fission. Dynamin and its binding protein amphiphysin cooperatively regulate membrane remodeling during the fission, but its precise mechanism remains elusive. In this study, we analyzed structural changes of dynamin-amphiphysin complexes during the membrane fission using electron microscopy (EM) and high-speed atomic force microscopy (HS-AFM)...
January 23, 2018: ELife
Gisel S Miszczuk, Ismael R Barosso, María Cecilia Larocca, Julieta Marrone, Raúl A Marinelli, Andrea C Boaglio, Enrique J Sánchez Pozzi, Marcelo G Roma, Fernando A Crocenzi
Impaired canalicular secretion due to increased endocytosis and intracellular retention of canalicular transporters such as BSEP and MRP2 is a main, common pathomechanism of cholestasis. Nevertheless, the mechanisms governing this process are unknown. We characterized this process in estradiol 17 β-d-glucuronide (E17G)-induced cholestasis, an experimental model which partially mimics pregnancy-induced cholestasis. Inhibition of clathrin-mediated endocytosis (CME) with monodansylcadaverine (MDC) or K+ depletion, but not the caveolin-mediated endocytosis inhibitors filipin and genistein, prevented E17G-induced endocytosis of BSEP and MRP2, and the associated impairment of activity of these transporters in isolated rat hepatocyte couplets (IRHC)...
January 17, 2018: Biochimica et Biophysica Acta
Azam S Alavi, Azadeh Meshkini
Although chemotherapy is an effective strategy for treatment of tumor cells, the non-specific distribution of chemotherapy drugs and their poor aqueous solubility result in impaired treatment and cause serious side effects in patients. In this study, mesoporous ZnO@Fe2O3 nanocomposite was fabricated and used as a platform for drug delivery. ZnO@Fe2O3 nanocomposite showed a high capacity for the adsorption of chemotherapy drug, methotrexate (MTX). Drug-loaded nanoparticle was coated with poly(ethylene glycol) (PEG), leading to the enhanced colloidal stability and good cytocompatibility...
January 15, 2018: European Journal of Pharmaceutical Sciences
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