Mammalian cell penetration, siRNA transfection, and DNA transfection by supercharged proteins.
AuthorsMcNaughton BR, Cronican JJ, Thompson DB, Liu DR,
JournalProc Natl Acad Sci U S A
PubMed ID19307578
'Nucleic acid reagents, including small interfering RNA (siRNA) and plasmid DNA, are important tools for the study of mammalian cells and are promising starting points for the development of new therapeutic agents. Realizing their full potential, however, requires nucleic acid delivery reagents that are simple to prepare, effective across many ... More
Glycosylation influences the lectin activities of the macrophage mannose receptor.
AuthorsSu Y, Bakker T, Harris J, Tsang C, Brown GD, Wormald MR, Gordon S, Dwek RA, Rudd PM, Martinez-Pomares L
JournalJ Biol Chem
PubMed ID15983039
'The mannose receptor (MR) is a heavily glycosylated endocytic receptor that recognizes both mannosylated and sulfated ligands through its C-type lectin domains and cysteine-rich (CR) domain, respectively. Differential binding properties have been described for MR isolated from different sources, and we hypothesized that this could be due to altered glycosylation. ... More
Caveolin regulates endocytosis of the muscle repair protein, dysferlin.
AuthorsHernández-Deviez DJ, Howes MT, Laval SH, Bushby K, Hancock JF, Parton RG,
JournalJ Biol Chem
PubMed ID18096699
'Dysferlin and Caveolin-3 are plasma membrane proteins associated with muscular dystrophy. Patients with mutations in the CAV3 gene show dysferlin mislocalization in muscle cells. By utilizing caveolin-null cells, expression of caveolin mutants, and different mutants of dysferlin, we have dissected the site of action of caveolin with respect to dysferlin ... More
AuthorsAustin CD, Lawrence DA, Peden AA, Varfolomeev EE, Totpal K, De Mazière AM, Klumperman J, Arnott D, Pham V, Scheller RH, Ashkenazi A
JournalProc Natl Acad Sci U S A
PubMed ID16801533
'Endocytosis is crucial for various aspects of cell homeostasis. Here, we show that proapoptotic death receptors (DRs) trigger selective destruction of the clathrin-dependent endocytosis machinery. DR stimulation induced rapid, caspase-mediated cleavage of key clathrin-pathway components, halting cellular uptake of the classic cargo protein transferrin. DR-proximal initiator caspases cleaved the clathrin ... More
Systems survey of endocytosis by multiparametric image analysis.
AuthorsCollinet C, Stöter M, Bradshaw CR, Samusik N, Rink JC, Kenski D, Habermann B, Buchholz F, Henschel R, Mueller MS, Nagel WE, Fava E, Kalaidzidis Y, Zerial M,
JournalNature
PubMed ID20190736
'Endocytosis is a complex process fulfilling many cellular and developmental functions. Understanding how it is regulated and integrated with other cellular processes requires a comprehensive analysis of its molecular constituents and general design principles. Here, we developed a new strategy to phenotypically profile the human genome with respect to transferrin ... More
VAMP4 cycles from the cell surface to the trans-Golgi network via sorting and recycling endosomes.
AuthorsTran TH, Zeng Q, Hong W
JournalJ Cell Sci
PubMed ID17327277
'VAMP4 is enriched in the trans-Golgi network (TGN) and functions in traffic from the early and recycling endosomes to the TGN, but its trafficking itinerary is unknown. Cells stably expressing TGN-enriched VAMP4 C-terminally-tagged with EGFP (VAMP4-EGFP) are able to internalize and transport EGFP antibody efficiently to the TGN, suggesting that ... More
Characterizing peptide-mediated DNA internalization in human cancer cells.
AuthorsWittrup A, Belting M,
JournalMethods Mol Biol
PubMed ID19085116
'Cell penetrating peptides (CPPs) are currently used to deliver various macromolecular cargos to intracellular sites of action both in vitro and in vivo on an experimental basis. During the last few years, even more evidence has accumulated indicating that the main route of entry for most CPPs is through endocytosis ... More
A burst of auxilin recruitment determines the onset of clathrin-coated vesicle uncoating.
AuthorsMassol RH, Boll W, Griffin AM, Kirchhausen T
JournalProc Natl Acad Sci U S A
PubMed ID16798879
'Clathrin-coated pits assemble on a membrane and pinch off as coated vesicles. The released vesicles then rapidly lose their clathrin coats in a process mediated by the ATPase Hsc70, recruited by auxilin, a J-domain-containing cofactor. How is the uncoating process regulated? We find that during coat assembly small and variable ... More
Src-like adaptor protein down-regulates T cell receptor (TCR)-CD3 expression by targeting TCRzeta for degradation.
AuthorsMyers MD, Dragone LL, Weiss A
JournalJ Cell Biol
PubMed ID16027224
'Src-like adaptor protein (SLAP) down-regulates expression of the T cell receptor (TCR)-CD3 complex during a specific stage of thymocyte development when the TCR repertoire is selected. Consequently, SLAP-/- thymocytes display alterations in thymocyte development. Here, we have studied the mechanism of SLAP function. We demonstrate that SLAP-deficient thymocytes have increased ... More
Fast, three-dimensional super-resolution imaging of live cells.
AuthorsJones SA, Shim SH, He J, Zhuang X,
JournalNat Methods
PubMed ID21552254
'We report super-resolution fluorescence imaging of live cells with high spatiotemporal resolution using stochastic optical reconstruction microscopy (STORM). By labeling proteins either directly or via SNAP tags with photoswitchable dyes, we obtained two-dimensional (2D) and 3D super-resolution images of living cells, using clathrin-coated pits and the transferrin cargo as model ... More
Ligands for clathrin-mediated endocytosis are differentially sorted into distinct populations of early endosomes.
AuthorsLakadamyali M, Rust MJ, Zhuang X
JournalCell
PubMed ID16530046
'Cells rely on the correct sorting of endocytic ligands and receptors for proper function. Early endosomes have been considered as the initial sorting station where cargos for degradation separate from those for recycling. Using live-cell imaging to monitor individual endosomes and ligand particles in real time, we have discovered a ... More
Regulation of CD4 expression via recycling by HRES-1/RAB4 controls susceptibility to HIV infection.
AuthorsNagy G, Ward J, Mosser DD, Koncz A, Gergely P, Stancato C, Qian Y, Fernandez D, Niland B, Grossman CE, Telarico T, Banki K, Perl A
JournalJ Biol Chem
PubMed ID16935861
'A novel 2986-base transcript encoded by the antisense strand of the HRES-1 human endogenous retrovirus was isolated from peripheral blood lymphocytes. This transcript codes for a 218-amino acid protein, termed HRES-1/Rab4, based on homology to the Rab4 family of small GTPases. Antibody 13407 raised against recombinant HRES-1/Rab4 detected a native ... More
Toll-like receptor 9-dependent activation by DNA-containing immune complexes is mediated by HMGB1 and RAGE.
AuthorsTian J, Avalos AM, Mao SY, Chen B, Senthil K, Wu H, Parroche P, Drabic S, Golenbock D, Sirois C, Hua J, An LL, Audoly L, La Rosa G, Bierhaus A, Naworth P, Marshak-Rothstein A, Crow MK, Fitzgerald KA, Latz E, Kiener PA, Coyle AJ
JournalNat Immunol
PubMed ID17417641
'Increased concentrations of DNA-containing immune complexes in the serum are associated with systemic autoimmune diseases such as lupus. Stimulation of Toll-like receptor 9 (TLR9) by DNA is important in the activation of plasmacytoid dendritic cells and B cells. Here we show that HMGB1, a nuclear DNA-binding protein released from necrotic ... More
Automated organelle-based colocalization in whole-cell imaging.
AuthorsWoodcroft BJ, Hammond L, Stow JL, Hamilton NA,
JournalCytometry A
PubMed ID19746416
The use of fluorescence microscopy to investigate protein colocalization is an invaluable tool for understanding subcellular structures and their associated proteins. However, current techniques are largely limited to two-dimensional (2D) imaging and often require manual segmentation. Here, we present OBCOL, a methodology to automatically segment and quantify protein colocalization not ... More
Copper-free click chemistry for dynamic in vivo imaging.
AuthorsBaskin JM, Prescher JA, Laughlin ST, Agard NJ, Chang PV, Miller IA, Lo A, Codelli JA, Bertozzi CR,
JournalProc Natl Acad Sci U S A
PubMed ID17942682
Dynamic imaging of proteins in live cells is routinely performed by using genetically encoded reporters, an approach that cannot be extended to other classes of biomolecules such as glycans and lipids. Here, we report a Cu-free variant of click chemistry that can label these biomolecules rapidly and selectively in living ... More
The utilization of pathogen-like cellular trafficking by single chain block copolymer.
Amphiphilic triblock copolymer, poly(ethylene oxide)-b-poly(propylene oxide)-b-poly(ethylene oxide), Pluronic P85, is unexpectedly shown to utilize sophisticated cellular trafficking mechanisms and enter brain microvessel endothelial cells and primary neurons that are poorly penetrable. Though caveolae serve as a primary entry site for the copolymer single chains, in cells devoid of caveolae, the ... More
Dynein is required for receptor sorting and the morphogenesis of early endosomes.
AuthorsDriskell OJ, Mironov A, Allan VJ, Woodman PG
JournalNat Cell Biol
PubMed ID17173037
The early endosome is organised into domains to ensure the separation of cargo. Activated mitogenic receptors, such as epidermal growth factor (EGF) receptor, are concentrated into vacuoles enriched for the small GTPase Rab5, which progressively exclude nutrient receptors, such as transferrin receptor, into neighbouring tubules. These vacuoles become enlarged, increase ... More
Molecular characterization of Rab11 interactions with members of the family of Rab11-interacting proteins.
AuthorsJunutula JR, Schonteich E, Wilson GM, Peden AA, Scheller RH, Prekeris R
JournalJ Biol Chem
PubMed ID15173169
The Rab11 subfamily of GTPases plays an important role in vesicle trafficking from endosomes to the plasma membrane. At least six Rab11 effectors (family of Rab11-interacting proteins (FIPs)) have been shown to interact with Rab11 and are hypothesized to regulate various membrane trafficking pathways such as transferrin recycling, cytokinesis, and ... More
Compartmental signal modulation: Endosomal phosphatidylinositol 3-phosphate controls endosome morphology and selective cargo sorting.
AuthorsFili N, Calleja V, Woscholski R, Parker PJ, Larijani B
JournalProc Natl Acad Sci U S A
PubMed ID17030795
It is increasingly recognized that the compartmental organization of signaling processes has a profound influence on cellular behavior. However, our inability to influence these compartmental events in a spatially restricted and acute manner limits our understanding of causation. To determine whether local compartmental loss of a phosphoinositide disrupts the normal ... More
Effects of Bafilomycin A1: an inhibitor of vacuolar H (+)-ATPases on endocytosis and apoptosis in RAW cells and RAW cell-derived osteoclasts.
AuthorsXu J, Feng HT, Wang C, Yip KH, Pavlos N, Papadimitriou JM, Wood D, Zheng MH
JournalJ Cell Biochem
PubMed ID12647307
Bafilomycin A1, a specific inhibitor of V-ATPases, is a potent inhibitor of bone resorption, but the underlying mechanisms of its action remain unclear. In this study, we investigated the effect of Bafilomycin A1 on endocytosis and apoptosis in RAW cells and RAW cell-derived osteoclasts. Quantitative analysis by flow cytometry showed ... More
Rab11b-mediated integrin recycling promotes brain metastatic adaptation and outgrowth.
Authors
JournalNat Commun
PubMed ID32541798
Rab5-regulated endocytosis plays a crucial role in apical extrusion of transformed cells.
Authors
JournalProc Natl Acad Sci U S A
PubMed ID28270608
Metabolic requirements of NK cells during the acute response against retroviral infection.
Authors
JournalNat Commun
PubMed ID34508086
HDAC6 mediates an aggresome-like mechanism for NLRP3 and pyrin inflammasome activation.
Authors
JournalScience
PubMed ID32943500
Cryo-EM of the dynamin polymer assembled on lipid membrane.
Authors
JournalNature
PubMed ID30069048
CD44 regulates epigenetic plasticity by mediating iron endocytosis.
Authors
JournalNat Chem
PubMed ID32747755
Tumor-Derived Extracellular Vesicles Breach the Intact Blood-Brain Barrier via Transcytosis.