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
Persistent cAMP-signals triggered by internalized G-protein-coupled receptors.
'G-protein-coupled receptors (GPCRs) are generally thought to signal to second messengers like cyclic AMP (cAMP) from the cell surface and to become internalized upon repeated or prolonged stimulation. Once internalized, they are supposed to stop signaling to second messengers but may trigger nonclassical signals such as mitogen-activated protein kinase (MAPK) ... More
Receptor and membrane recycling can occur with unaltered efficiency despite dramatic Rab5(q79l)-induced changes in endosome geometry.
AuthorsCeresa BP, Lotscher M, Schmid SL
JournalJ Biol Chem
PubMed ID11136733
'Current models for sorting in the endosomal compartment suggest that endosomal geometry plays a significant role as membrane-bound proteins accumulate in tubular regions for recycling, and lumenal markers accumulate in large vacuolar portions for delivery to lysosomes. Rab5, a small molecular weight GTPase, functions in the formation and maintenance of ... More
SLAM is a microbial sensor that regulates bacterial phagosome functions in macrophages.
AuthorsBerger SB, Romero X, Ma C, Wang G, Faubion WA, Liao G, Compeer E, Keszei M, Rameh L, Wang N, Boes M, Regueiro JR, Reinecker HC, Terhorst C,
JournalNat Immunol
PubMed ID20818396
'Phagocytosis is a pivotal process by which macrophages eliminate microorganisms after recognition by pathogen sensors. Here we unexpectedly found that the self ligand and cell surface receptor SLAM functioned not only as a costimulatory molecule but also as a microbial sensor that controlled the killing of gram-negative bacteria by macrophages. ... More
The small GTPase Rab22 interacts with EEA1 and controls endosomal membrane trafficking.
AuthorsKauppi M, Simonsen A, Bremnes B, Vieira A, Callaghan J, Stenmark H, Olkkonen VM
JournalJ Cell Sci
PubMed ID11870209
'Rab22a is a small GTPase that is expressed ubiquitously in mammalian tissues and displays the highest sequence homology to Rab5. In BHK-21 cells, overexpression of the wild-type Rab22a caused formation of abnormally large vacuole-like structures containing the early-endosomal antigen EEA1 but not Rab11, a marker of recycling endosomes or the ... More
Regulation of caveolar endocytosis by syntaxin 6-dependent delivery of membrane components to the cell surface.
AuthorsChoudhury A, Marks DL, Proctor KM, Gould GW, Pagano RE
JournalNat Cell Biol
PubMed ID16565709
'Caveolar endocytosis has an important function in the cellular uptake of some bacterial toxins, viruses and circulating proteins. However, the molecular machinery involved in regulating caveolar uptake is poorly defined. Here, we demonstrate that caveolar endocytosis is regulated by syntaxin 6, a target membrane soluble N-ethylmaleimide attachment protein receptor (t-SNARE) ... More
The Ankrd 13 family of UIM-bearing proteins regulates EGF receptor endocytosis from the plasma membrane.
AuthorsTanno H, Yamaguchi T, Goto E, Ishido S, Komada M,
JournalMol Biol Cell
PubMed ID22298428
'The mechanism of ubiquitin-dependent endocytosis of cell surface proteins is not completely understood. Here we examine the role of the ankyrin repeat domain (Ankrd) 13A, 13B, and 13D proteins, which constitute a functionally unknown family of ubiquitin-interacting motif (UIM)-bearing proteins, in the process. Stimulation of human HeLa cells with epidermal ... More
Rapidly inducible changes in phosphatidylinositol 4,5-bisphosphate levels influence multiple regulatory functions of the lipid in intact living cells.
AuthorsVarnai P, Thyagarajan B, Rohacs T, Balla T
JournalJ Cell Biol
PubMed ID17088424
'Rapamycin (rapa)-induced heterodimerization of the FRB domain of the mammalian target of rapa and FKBP12 was used to translocate a phosphoinositide 5-phosphatase (5-ptase) enzyme to the plasma membrane (PM) to evoke rapid changes in phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P(2)) levels. Rapa-induced PM recruitment of a truncated type IV 5-ptase containing only the ... More
EphrinB-EphB signalling regulates clathrin-mediated endocytosis through tyrosine phosphorylation of synaptojanin 1.
AuthorsIrie F, Okuno M, Pasquale EB, Yamaguchi Y
JournalNat Cell Biol
PubMed ID15821731
'Recent studies show that Eph receptors act mainly through the regulation of actin reorganization. Here, we show a novel mode of action for EphB receptors. We identify synaptojanin 1 - a phosphatidylinositol 5''-phosphatase that is involved in clathrin-mediated endocytosis - as a physiological substrate for EphB2. EphB2 causes tyrosine phosphorylation ... More
Rab15 mediates an early endocytic event in Chinese hamster ovary cells.
AuthorsZuk PA, Elferink LA
JournalJ Biol Chem
PubMed ID10428799
'Rab GTPases comprise a large family of monomeric proteins that regulate a diverse number of membrane trafficking events, including endocytosis. In this paper, we examine the subcellular distribution and function of the GTPase Rab15. Our biochemical and confocal immunofluorescence studies demonstrate that Rab15 associates with the transferrin receptor, a marker ... More
Identification of syntenin as a protein of the apical early endocytic compartment in Madin-Darby canine kidney cells.
AuthorsFialka I, Steinlein P, Ahorn H, Böck G, Burbelo PD, Haberfellner M, Lottspeich F, Paiha K, Pasquali C, Huber LA
JournalJ Biol Chem
PubMed ID10473577
'We used flow cytometry to sort and analyze apical and basolateral endocytic vesicles from filter-grown Madin-Darby canine kidney (MDCK) cells after membrane internalization of the lipophilic fluorescent probe trimethylamino-diphenylhexatriene. Western blot analysis of sorted fractions showed enrichment of the early endosomal markers transferrin receptor and the small GTPase Rab5. Two-dimensional ... More
Trafficking defects in WASH-knockout fibroblasts originate from collapsed endosomal and lysosomal networks.
The Arp2/3-activator Wiskott-Aldrich syndrome protein and Scar homologue (WASH) is suggested to regulate actin-dependent membrane scission during endosomal sorting, but its cellular roles have not been fully elucidated. To investigate WASH function, we generated tamoxifen-inducible WASH-knockout mouse embryonic fibroblasts (WASHout MEFs). Of interest, although EEA1(+) endosomes were enlarged, collapsed, and ... More
An eight residue fragment of an acyl carrier protein suffices for post-translational introduction of fluorescent pantetheinyl arms in protein modification in vitro and in vivo.
AuthorsZhou Z, Koglin A, Wang Y, McMahon AP, Walsh CT,
JournalJ Am Chem Soc
PubMed ID18593165
Genetically encoded tags for tracking a given protein continue to be of great interest in a multitude of in vitro and in vivo contexts. Acyl carrier proteins, both free-standing and as embedded 80-100 residue domains, contain a specific serine side chain that undergoes post-translational pantetheinylation from CoASH as donor substrate. ... More
C-peptide is internalised in human endothelial and vascular smooth muscle cells via early endosomes.
AuthorsLuppi P, Geng X, Cifarelli V, Drain P, Trucco M,
JournalDiabetologia
PubMed ID19662378
AIMS/HYPOTHESIS: There is increasing evidence that C-peptide exerts intracellular effects in a variety of cells and could be beneficial in patients with type 1 diabetes. Exactly how C-peptide achieves these effects, however, is unknown. Recent reports showed that C-peptide internalised in the cytoplasm of HEK-293 and Swiss 3T3 cells, where ... 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
Nicotinic acetylcholine receptor is internalized via a Rac-dependent, dynamin-independent endocytic pathway.
AuthorsKumari S, Borroni V, Chaudhry A, Chanda B, Massol R, Mayor S, Barrantes FJ,
JournalJ Cell Biol
PubMed ID18591431
Endocytosis of the nicotinic acetylcholine receptor (AChR) is a proposed major mechanism of neuromodulation at neuromuscular junctions and in the pathology of synapses in the central nervous system. We show that binding of the competitive antagonist alpha-bungarotoxin (alphaBTX) or antibody-mediated cross-linking induces the internalization of cell surface AChR to late ... More
Evidence to support the cellular mechanism involved in serum IgG homeostasis in humans.
AuthorsWard ES, Zhou J, Ghetie V, Ober RJ
JournalInt Immunol
PubMed ID12578848
IgG is the most abundant serum antibody and is an essential component of the humoral immune response. It is known that the 'neonatal' Fc receptor (FcRn) plays a role in maintaining constant serum IgG levels by acting as a protective receptor which binds and salvages IgG from degradation. However, the ... More
Perforin triggers a plasma membrane-repair response that facilitates CTL induction of apoptosis.
Perforin delivers granzymes to induce target-cell apoptosis. At high concentrations, perforin multimerizes in the plasma membrane to form pores. However, whether granzymes enter target cells via membrane pores is uncertain. Here we find that perforin at physiologically relevant concentrations and during cell-mediated lysis creates pores in the target-cell membrane, transiently ... More
Endosomal recycling controls plasma membrane area during mitosis.
AuthorsBoucrot E, Kirchhausen T
JournalProc Natl Acad Sci U S A
PubMed ID17483462
The shape and total surface of a cell and its daughters change during mitosis. Many cells round up during prophase and metaphase and reacquire their extended and flattened shape during cytokinesis. How does the total area of plasma membrane change to accommodate these morphological changes and by what mechanism is ... More
Endocytosed transferrin receptors recycle via distinct dynamin and phosphatidylinositol 3-kinase-dependent pathways.
Authorsvan Dam EM, Ten Broeke T, Jansen K, Spijkers P, Stoorvogel W
JournalJ Biol Chem
PubMed ID12372835
Recycling of endocytosed membrane proteins involves passage through early endosomes and recycling endosomes. Previously, we demonstrated a role for clathrin-coated vesicles in transferrin receptor recycling. These clathrin-coated vesicles are formed from recycling endosomes in a process that was inhibited in dynamin-1(G273D)-overexpressing cells. Here we show a second transferrin recycling pathway, ... More
T cell receptor ligation induces the formation of dynamically regulated signaling assemblies.
AuthorsBunnell SC, Hong DI, Kardon JR, Yamazaki T, McGlade CJ, Barr VA, Samelson LE
JournalJ Cell Biol
PubMed ID12356870
Tcell antigen receptor (TCR) ligation initiates tyrosine kinase activation, signaling complex assembly, and immune synapse formation. Here, we studied the kinetics and mechanics of signaling complex formation in live Jurkat leukemic T cells using signaling proteins fluorescently tagged with variants of enhanced GFP (EGFP). Within seconds of contacting coverslips coated ... More
Uptake and intracellular transport of acidic fibroblast growth factor: evidence for free and cytoskeleton-anchored fibroblast growth factor receptors.
AuthorsCitores L, Wesche J, Kolpakova E, Olsnes S
JournalMol Biol Cell
PubMed ID10564275
Endocytic uptake and intracellular transport of acidic FGF was studied in cells transfected with FGF receptor 4 (FGFR4). Acidification of the cytosol to block endocytic uptake from coated pits did not inhibit endocytosis of the growth factor in COS cells transfected with FGFR4, indicating that it is to a large ... More
Transferrin receptor recycling in the absence of perinuclear recycling endosomes.
AuthorsSheff D, Pelletier L, O'Connell CB, Warren G, Mellman I
JournalJ Cell Biol
PubMed ID11877458
In mammalian cells, internalized receptors such as transferrin (Tfn) receptor are presumed to pass sequentially through early endosomes (EEs) and perinuclear recycling endosomes (REs) before returning to the plasma membrane. Whether passage through RE is obligatory, however, remains unclear. Kinetic analysis of endocytosis in CHO cells suggested that the majority ... More
Mechanism of uptake of C105Y, a novel cell-penetrating peptide.
AuthorsRhee M, Davis P
JournalJ Biol Chem
PubMed ID16272160
C105Y, a synthetic peptide (CSIPPEVKFNKPFVYLI) based on the amino acid sequence corresponding to residues 359-374 of alpha1-antitrypsin, enhances gene expression from DNA nanoparticles. To investigate how this enhancement occurs, C105Y was fluorescently labeled to study its uptake and intracellular trafficking. When human hepatoma cells (HuH7) were incubated with fluorescently labeled ... More
Endocytic recycling compartments altered in cisplatin-resistant cancer cells.
AuthorsLiang XJ, Mukherjee S, Shen DW, Maxfield FR, Gottesman MM
JournalCancer Res
PubMed ID16489040
The clinical utility of cisplatin to treat human malignancies is often limited by the development of drug resistance. We have previously shown that cisplatin-resistant human KB adenocarcinoma cells that are cross-resistant to methotrexate and heavy metals have altered endocytic recycling. In this work, we tracked lipids in the endocytic recycling ... More
Dominant-interfering Hsc70 mutants disrupt multiple stages of the clathrin-coated vesicle cycle in vivo.
AuthorsNewmyer SL, Schmid SL
JournalJ Cell Biol
PubMed ID11157986
Within the clathrin-coated vesicle (CCV) cycle, coat assembly drives the internalization of receptors from the cell surface and disassembly allows for the processing of internalized ligands. The heat shock cognate protein, hsc70, has been implicated in regulating coat disassembly. We find that in cells overexpressing ATPase-deficient hsc70 mutants, uncoating of ... More
Role of phosphatidic acid phosphatase 2a in uptake of extracellular lipid phosphate mediators.
AuthorsRoberts RZ, Morris AJ
JournalBiochim Biophys Acta
PubMed ID10962286
Phosphatidic acid (PA) phosphatase 2a (PAP2a) is an integral membrane glycoprotein that hydrolyzes a number of structurally related lipid phosphate substrates when presented in mixed phospholipid and detergent micelles. The physiological substrate specificity and functions of this enzyme are unclear. Using reconstitution studies we demonstrate that PAP2a hydrolyses both PA ... More
Adenovirus triggers macropinocytosis and endosomal leakage together with its clathrin-mediated uptake.
Adenovirus type 2 (Ad2) binds the coxsackie B virus Ad receptor and is endocytosed upon activation of the alphav integrin coreceptors. Here, we demonstrate that expression of dominant negative clathrin hub, eps15, or K44A-dynamin (dyn) inhibited Ad2 uptake into epithelial cells, indicating clathrin-dependent viral endocytosis. Surprisingly, Ad strongly stimulated the ... More
Alkyl-lysophospholipid accumulates in lipid rafts and induces apoptosis via raft-dependent endocytosis and inhibition of phosphatidylcholine synthesis.
Authorsvan der Luit AH, Budde M, Ruurs P, Verheij M, van Blitterswijk WJ
JournalJ Biol Chem
PubMed ID12183451
The synthetic alkyl-lysophospholipid (ALP), 1-O-octadecyl-2-O-methyl-rac-glycero-3-phosphocholine, is an antitumor agent that acts on cell membranes and can induce apoptosis. We investigated how ALP is taken up by cells, how it affects de novo biosynthesis of phosphatidylcholine (PC), and how critical this is to initiate apoptosis. We compared an ALP-sensitive mouse lymphoma ... More
G protein-coupled receptor endocytosis in ADP-ribosylation factor 6-depleted cells.
AuthorsHoundolo T, Boulay PL, Claing A
JournalJ Biol Chem
PubMed ID15590645
The internalization of G protein-coupled receptors is regulated by several important proteins that act in concert to finely control this complex cellular process. Here, we have applied the RNA interference approach to demonstrate that ADP-ribosylation factor 6 (ARF6) is essential for the endocytosis of a broad variety of receptors. Reduction ... More
Surface functionalization of doxorubicin-loaded liposomes with octa-arginine for enhanced anticancer activity.
Doxorubicin-loaded PEGylated liposomes (commercially available as DOXIL or Lipodox) were surface functionalized with a cell-penetrating peptide, octa-arginine (R8). For this purpose, R8-peptide was conjugated to the polyethylene glycol-dioleoyl phosphatidylethanolamine (PEG-DOPE) amphiphilic co-polymer. The resultant R8-PEG-PE conjugate was introduced into the lipid bilayer of liposomes at 2 mol% of total lipid ... More
The reversion-inducing cysteine-rich protein with Kazal motifs (RECK) interacts with membrane type 1 matrix metalloproteinase and CD13/aminopeptidase N and modulates their endocytic pathways.
AuthorsMiki T, Takegami Y, Okawa K, Muraguchi T, Noda M, Takahashi C
JournalJ Biol Chem
PubMed ID17329256
The reversion-inducing cysteine-rich protein with Kazal motifs (RECK) is anchored to the cell surface via glycosylphosphatidylinositol. This molecule antagonizes the function of membrane type 1 matrix metalloproteinase (MT1-MMP) to promote proMMP-2 maturation. Here, we attempt to clarify the mechanism underlying RECK functions. First, we found that RECK forms a complex ... More
ClC-3 chloride channels facilitate endosomal acidification and chloride accumulation.
AuthorsHara-Chikuma M, Yang B, Sonawane ND, Sasaki S, Uchida S, Verkman AS
JournalJ Biol Chem
PubMed ID15504734
We investigated the involvement of ClC-3 chloride channels in endosomal acidification by measurement of endosomal pH and chloride concentration [Cl-] in control versus ClC-3-deficient hepatocytes and in control versus ClC-3-transfected Chinese hamster ovary cells. Endosomes were labeled with pH or [Cl-]-sensing fluorescent transferrin (Tf), which targets to early/recycling endosomes, or ... More
Dissecting the heterogeneity of DENV vaccine-elicited cellular immunity using single-cell RNA sequencing and metabolic profiling.
Authors
JournalNat Commun
PubMed ID31413301
A single genetic locus controls both expression of DPEP1/CHMP1A and kidney disease development via ferroptosis.
Authors
JournalNat Commun
PubMed ID34426578
Brain Delivery of Multifunctional Dendrimer Protein Bioconjugates.
Authors
JournalAdv Sci (Weinh)
PubMed ID29876217
Lung emphysema and impaired macrophage elastase clearance in mucolipin 3 deficient mice.
Authors
JournalNat Commun
PubMed ID35031603
Dynamin regulates the dynamics and mechanical strength of the actin cytoskeleton as a multifilament actin-bundling protein.
Authors
JournalNat Cell Biol
PubMed ID32451441
Endophilin-A3 and Galectin-8 control the clathrin-independent endocytosis of CD166.
Authors
JournalNat Commun
PubMed ID32193381
Visualization of self-delivering hydrophobically modified siRNA cellular internalization.
Authors
JournalNucleic Acids Res
PubMed ID27899655
SNAREs define targeting specificity of trafficking vesicles by combinatorial interaction with tethering factors.
Authors
JournalNat Commun
PubMed ID30962439
A fluorescence-based in vitro assay for investigating early endosome dynamics.