Protection of mammalian cells from diphtheria toxin by exogenous nucleotides

1979 ◽  
Vol 25 (3) ◽  
pp. 285-290 ◽  
Author(s):  
John L. Middlebrook ◽  
Rebecca B. Dorland
Keyword(s):  
Diphtheria Toxin ◽  
Mammalian Cells ◽  
Cell Surface Receptor ◽  
Lethal Effects ◽  
Cellular Protection ◽  
Future Studies ◽  
Degree Of Protection ◽  
Phosphate Chain ◽  
Surface Receptor ◽  
Toxin Receptor

Exogenous nucleotides were found to protect mammalian cells from the lethal effects of diphtheria toxin. Protective potency of a given nucleotide was base specific and phosphate chain length dependent. Full expression of protective potency required an intact nucleotide, but the effect did not appear to be mediated by nucleotide-induced phosphorylation. Nucleotides antagonized the binding of diphtheria toxin to its cell surface receptor in a manner that correlated with the degree of protection. It was concluded that cellular protection from diphtheria toxin by nucleotides results from inhibition of toxin–receptor binding and that nucleotides therefore may serve as valuable research tools for future studies.

scholarly journals Endocytic Delivery of Vancomycin Mediated by a Synthetic Cell Surface Receptor:  Rescue of Bacterially Infected Mammalian Cells and Tissue Targeting In Vivo

2007 ◽  
Vol 129 (2) ◽  
pp. 268-269 ◽  
Author(s):  
Siwarutt Boonyarattanakalin ◽  
Jianfang Hu ◽  
Sheryl A. Dykstra-Rummel ◽  
Avery August ◽  
Blake R. Peterson
Keyword(s):  
Cell Surface ◽  
Mammalian Cells ◽  
Cell Surface Receptor ◽  
Synthetic Cell ◽  
Tissue Targeting ◽  
Surface Receptor

scholarly journals Neuropilin-1 and heparan sulfate proteoglycans cooperate in cellular uptake of nanoparticles functionalized by cationic cell-penetrating peptides

2015 ◽  
Vol 1 (10) ◽  
pp. e1500821 ◽  
Author(s):  
Hong-Bo Pang ◽  
Gary B. Braun ◽  
Erkki Ruoslahti
Keyword(s):  
Cell Surface ◽  
Heparan Sulfate ◽  
Vascular Permeability ◽  
Mammalian Cells ◽  
Cell Penetrating Peptides ◽  
Proteolytic Processing ◽  
Cell Surface Receptor ◽  
Neuropilin 1 ◽  
Cell Penetrating ◽  
Surface Receptor

Cell-penetrating peptides (CPPs) have been widely used to deliver nanomaterials and other types of macromolecules into mammalian cells for therapeutic and diagnostic use. Cationic CPPs that bind to heparan sulfate (HS) proteoglycans on the cell surface induce potent endocytosis; however, the role of other surface receptors in this process is unclear. We describe the convergence of an HS-dependent pathway with the C-end rule (CendR) mechanism that enables peptide ligation with neuropilin-1 (NRP1), a cell surface receptor known to be involved in angiogenesis and vascular permeability. NRP1 binds peptides carrying a positive residue at the carboxyl terminus, a feature that is compatible with cationic CPPs, either intact or after proteolytic processing. We used CPP and CendR peptides, as well as HS- and NRP1-binding motifs from semaphorins, to explore the commonalities and differences of the HS and NRP1 pathways. We show that the CendR-NRP1 interaction determines the ability of CPPs to induce vascular permeability. We also show at the ultrastructural level, using a novel cell entry synchronization method, that both the HS and NRP1 pathways can initiate a macropinocytosis-like process and visualize these CPP-cargo complexes going through various endosomal compartments. Our results provide new insights into how CPPs exploit multiple surface receptor pathways for intracellular delivery.

scholarly journals Specific Capture of Mammalian Cells by Cell Surface Receptor Binding to Ligand Immobilized on Gold Thin Films

2006 ◽  
Vol 5 (7) ◽  
pp. 1580-1585 ◽  
Author(s):  
Dora Peelen ◽  
Voula Kodoyianni ◽  
Jieun Lee ◽  
Ting Zheng ◽  
Michael R. Shortreed ◽  
...  
Keyword(s):  
Thin Films ◽  
Cell Surface ◽  
Receptor Binding ◽  
Mammalian Cells ◽  
Cell Surface Receptor ◽  
Surface Receptor ◽  
Gold Thin Films

scholarly journals Tissue distribution of mRNA for heparin-binding epidermal growth factor

1992 ◽  
Vol 287 (3) ◽  
pp. 681-684 ◽  
Author(s):  
T J Vaughan ◽  
J C Pascall ◽  
K D Brown
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Diphtheria Toxin ◽  
Cell Surface Receptor ◽  
Heparin Binding ◽  
Reverse Transcription Pcr ◽  
Body Tissues ◽  
Wide Range ◽  
Surface Receptor ◽  
Epidermal Growth

Heparin-binding epidermal growth factor (HB-EGF) is a recently identified member of the EGF family. Mature HB-EGF is processed from a larger transmembrane precursor which can itself act as a cell-surface receptor for the internalization of diphtheria toxin into eukaryotic cells. However, to date there is no information available on the distribution of HB-EGF in mammalian tissues. We have therefore used reverse-transcription PCR to analyse the expression of HB-EGF mRNA in a wide range of tissues. HB-EGF transcripts were detected in RNA isolated from 15 of the 22 tissues obtained from adult pigs, which is consistent with the ability of diphtheria toxin to affect many body tissues.

scholarly journals Subcellular Redistribution of Pit-2 PiTransporter/Amphotropic Leukemia Virus (A-MuLV) Receptor in A-MuLV-Infected NIH 3T3 Fibroblasts: Involvement in Superinfection Interference

2000 ◽  
Vol 74 (6) ◽  
pp. 2847-2854 ◽  
Author(s):  
Zsolt Jobbagy ◽  
Susan Garfield ◽  
Lisa Baptiste ◽  
Maribeth V. Eiden ◽  
Wayne B. Anderson
Keyword(s):  
Plasma Membrane ◽  
Mammalian Cells ◽  
Leukemia Virus ◽  
Cell Surface Receptor ◽  
3T3 Cells ◽  
Sodium Dependent ◽  
Surface Receptor ◽  
A Cell ◽  
Nih 3T3 ◽  
Nih 3T3 Cells

ABSTRACT Amphotropic murine leukemia virus (A-MuLV) utilizes the Pit-2 sodium-dependent phosphate transporter as a cell surface receptor to infect mammalian cells. Previous studies established that infection of cells with A-MuLV resulted in the specific down-modulation of phosphate uptake mediated by Pit-2 and in resistance to superinfection with A-MuLV. To study the mechanisms underlying these phenomena, we constructed plasmids capable of efficiently expressing ɛ epitope- and green fluorescent protein (GFP)-tagged human Pit-2 proteins in mammalian cells. Overexpression of ɛ-epitope-tagged Pit-2 transporters in NIH 3T3 cells resulted in a marked increase in sodium-dependent Pi uptake. This increase in Piuptake was specifically blocked by A-MuLV infection but not by infection with ecotropic MuLV (E-MuLV) (which utilizes a cationic amino acid transporter, not Pit-2, as a cell surface receptor). These data, together with the finding that the tagged Pit-2 transporters retained their A-MuLV receptor function, indicate that the insertion of epitope tags does not affect either retrovirus receptor or Pitransporter function. The overexpressed epitope-tagged transporters were detected in cell lysates, by Western blot analysis using both ɛ-epitope- and GFP-specific antibodies as well as with Pit-2 antiserum. Both the epitope- and GFP-tagged transporters showed almost exclusive plasma membrane localization when expressed in NIH 3T3 cells, as determined by laser scanning confocal microscopy. Importantly, when NIH 3T3 cells expressing these proteins were productively infected with A-MuLV, the tagged transporters and receptors were no longer detected in the plasma membrane but rather were localized to a punctate structure within the cytosolic compartment distinct from Golgi, endoplasmic reticulum, endosomes, lysosomes, and mitochondria. The intracellular Pit-2 pool colocalized with the virus in A-MuLV-infected cells. A similar redistribution of the tagged Pit-2 proteins was not observed following infection with E-MuLV, indicating that the redistribution of Pit-2 is not directly attributable to general effects associated with retroviral infection but rather is a specific consequence of A-MuLV–Pit-2 interactions.

scholarly journals The p21 Rho-activating Toxin Cytotoxic Necrotizing Factor 1 Is Endocytosed by a Clathrin-independent Mechanism and Enters the Cytosol by an Acidic-dependent Membrane Translocation Step

2000 ◽  
Vol 11 (5) ◽  
pp. 1775-1787 ◽  
Author(s):  
Stephanette Contamin ◽  
Antoine Galmiche ◽  
Anne Doye ◽  
Gilles Flatau ◽  
Alexandre Benmerah ◽  
...  
Keyword(s):  
Diphtheria Toxin ◽  
Catalytic Domain ◽  
Dominant Negative ◽  
Cell Surface Receptor ◽  
Bacterial Toxin ◽  
Membrane Translocation ◽  
Plant Toxin ◽  
Cytotoxic Necrotizing Factor ◽  
Surface Receptor ◽  
Cytotoxic Necrotizing Factor 1

Cytotoxic necrotizing factor 1 (CNF1), a protein produced by pathogenic strains of Escherichia coli, activates the p21 Rho-GTP-binding protein, inducing a profound reorganization of the actin cytoskeleton. CNF1 binds to its cell surface receptor on HEp-2 cells with high affinity (Kd= 20 pM). In HEp-2 cells the action of CNF1 is not blocked in the presence of filipin, a drug described to reduce cholera toxin internalization by the caveolae-like mechanism. Moreover, HEp-2 cells, which express a dominant negative form of proteins that impair the formation of clathrin coated-vesicles and internalization of transferrin (Eps15, dynamin or intersectin-Src homology 3), are still sensitive to CNF1. In this respect, the endocytosis of CNF1 is similar to the plant toxin ricin. However, unlike ricin toxin, CNF1 does not cross the Golgi apparatus and requires an acidic cell compartment to transfer its enzymatic activity into the cytosol in a manner similar to that required by diphtheria toxin. As shown for diphtheria toxin, the pH-dependent membrane translocation step of CNF1 could be mimicked at the level of the plasma membrane by a brief exposure to a pH of ≤5.2. CNF1 is the first bacterial toxin described that uses both a clathrin-independent endocytic mechanism and an acidic-dependent membrane translocation step in its delivery of the catalytic domain to the cell cytosol.

Endocytosis

1997 ◽  
Vol 77 (3) ◽  
pp. 759-803 ◽  
Author(s):  
S. Mukherjee ◽  
R. N. Ghosh ◽  
F. R. Maxfield
Keyword(s):  
Antigen Presentation ◽  
Mammalian Cells ◽  
Membrane Lipids ◽  
Extracellular Fluid ◽  
Cell Polarization ◽  
Receptor Expression ◽  
Cell Surface Receptor ◽  
Coated Pits ◽  
Surface Receptor ◽  
Membrane Components

Mammalian cells take up extracellular material by a variety of different mechanisms that are collectively termed endocytosis. Endocytic mechanisms serve many important cellular functions including the uptake of extracellular nutrients, regulation of cell-surface receptor expression, maintenance of cell polarity, and antigen presentation. Endocytic pathways are also utilized by viruses, toxins, and symbiotic microorganisms to gain entry into cells. One of the best-characterized endocytic mechanisms is receptor-mediated endocytosis via clathrin-coated pits. This type of endocytosis constitutes the major emphasis of this review, with a brief discussion of other endocytic mechanisms and their comparison with the receptor-mediated pathway. This review describes and evaluates critically current understanding of the mechanisms of entry of plasma membrane components such as the receptor-ligand complexes and membrane lipids as well as the extracellular fluid into cells. The intracellular sorting and trafficking of these molecules upon internalization are also described. The roles of endocytosis in physiological and pathological processes are discussed. These include maintenance of cell polarization, antigen presentation, glucose transport, atherosclerosis, Alzheimer's disease, and the endocytosis of toxins and viruses.

scholarly journals Anthrax toxin triggers endocytosis of its receptor via a lipid raft–mediated clathrin-dependent process

2003 ◽  
Vol 160 (3) ◽  
pp. 321-328 ◽  
Author(s):  
Laurence Abrami ◽  
Shihui Liu ◽  
Pierre Cosson ◽  
Stephen H. Leppla ◽  
F. Gisou van der Goot
Keyword(s):  
Lipid Rafts ◽  
Protective Antigen ◽  
Lethal Factor ◽  
Physiological Role ◽  
Membrane Lipid ◽  
Anthrax Toxin ◽  
Cell Surface Receptor ◽  
Surface Receptor ◽  
Toxin Receptor ◽  
Major Route

The protective antigen (PA) of the anthrax toxin binds to a cell surface receptor and thereby allows lethal factor (LF) to be taken up and exert its toxic effect in the cytoplasm. Here, we report that clustering of the anthrax toxin receptor (ATR) with heptameric PA or with an antibody sandwich causes its association to specialized cholesterol and glycosphingolipid-rich microdomains of the plasma membrane (lipid rafts). We find that although endocytosis of ATR is slow, clustering it into rafts either via PA heptamerization or using an antibody sandwich is necessary and sufficient to trigger efficient internalization and allow delivery of LF to the cytoplasm. Importantly, altering raft integrity using drugs prevented LF delivery and cleavage of cytosolic MAPK kinases, suggesting that lipid rafts could be therapeutic targets for drugs against anthrax. Moreover, we show that internalization of PA is dynamin and Eps15 dependent, indicating that the clathrin-dependent pathway is the major route of anthrax toxin entry into the cell. The present work illustrates that although the physiological role of the ATR is unknown, its trafficking properties, i.e., slow endocytosis as a monomer and rapid clathrin-mediated uptake on clustering, make it an ideal anthrax toxin receptor.

scholarly journals Identification of diphtheria toxin receptor and a nonproteinous diphtheria toxin-binding molecule in Vero cell membrane.

1988 ◽  
Vol 107 (2) ◽  
pp. 511-519 ◽  
Author(s):  
E Mekada ◽  
Y Okada ◽  
T Uchida
Keyword(s):  
Cell Membrane ◽  
Vero Cell ◽  
Diphtheria Toxin ◽  
Vero Cells ◽  
Binding Activity ◽  
Membrane Preparation ◽  
Toxin Binding ◽  
Surface Receptor ◽  
Toxin Receptor ◽  
Cell Membrane Preparation

Two substances possessing the ability to bind to diphtheria toxin (DT) were found to be present in a membrane fraction from DT-sensitive Vero cells. One of these substances was found on the basis of its ability to bind DT and inhibit its cytotoxic effect. This inhibitory substance competitively inhibited the binding of DT to Vero cells. However this inhibitor could not bind to CRM197, the product of a missense mutation in the DT gene, and did not inhibit the binding of CRM197 to Vero cells. Moreover, similar levels of the inhibitory activity were observed in membrane fractions from DT-insensitive mouse cells, suggesting the inhibitor is not the DT receptor which is specifically present in DT-sensitive cells. The second DT-binding substance was found in the same Vero cell membrane preparation by assaying the binding of 125I-labeled CRM197. Such DT-binding activity could not be observed in membrane preparation from mouse L cells. From competition studies using labeled DT and CRM proteins, we conclude that this binding activity is due to the surface receptor for DT. Treatment of these substances with several enzymes revealed that the inhibitor was sensitive to certain RNases but resistant to proteases, whereas the DT receptor was resistant to RNase but sensitive to proteases. The receptor was solubilized and partially purified by chromatography on CM-Sepharose column. Immunoprecipitation and Western blotting analysis of the partially purified receptor revealed that a 14.5-kD protein is the DT receptor, or at least a component of it.

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