scholarly journals Interactions between diphtheria toxin entry and anion transport in vero cells. III. Effect on toxin binding and anion transport of tumor-promoting phorbol esters, vanadate, fluoride, and salicylate.

1986 ◽  
Vol 261 (4) ◽  
pp. 1562-1569 ◽  
Author(s):  
S Olsnes ◽  
E Carvajal ◽  
K Sandvig
Keyword(s):  
Diphtheria Toxin ◽  
Phorbol Esters ◽  
Vero Cells ◽  
Anion Transport ◽  
Toxin Binding

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.

scholarly journals Association of diphtheria toxin with Vero cells. Demonstration of a receptor.

1978 ◽  
Vol 253 (20) ◽  
pp. 7325-7330
Author(s):  
J.L. Middlebrook ◽  
R.B. Dorland ◽  
S.H. Leppla
Keyword(s):  
Diphtheria Toxin ◽  
Vero Cells

Anti-human HB-EGF monoclonal antibodies inhibiting ectodomain shedding of HB-EGF and diphtheria toxin binding

2010 ◽  
Vol 148 (1) ◽  
pp. 55-69 ◽  
Author(s):  
M. Hamaoka ◽  
I. Chinen ◽  
T. Murata ◽  
S. Takashima ◽  
R. Iwamoto ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Diphtheria Toxin ◽  
Ectodomain Shedding ◽  
Toxin Binding

scholarly journals The 27-kD diphtheria toxin receptor-associated protein (DRAP27) from vero cells is the monkey homologue of human CD9 antigen: expression of DRAP27 elevates the number of diphtheria toxin receptors on toxin-sensitive cells.

1992 ◽  
Vol 118 (6) ◽  
pp. 1389-1399 ◽  
Author(s):  
T Mitamura ◽  
R Iwamoto ◽  
T Umata ◽  
T Yomo ◽  
I Urabe ◽  
...  
Keyword(s):  
Diphtheria Toxin ◽  
Binding Capacity ◽  
Hybrid Cell ◽  
Antigen Expression ◽  
Vero Cells ◽  
Mouse Cell ◽  
Homology Search ◽  
Receptor Associated Protein ◽  
New Family ◽  
Cloned Cdna

Diphtheria toxin (DT) receptor associates with a 27-kD membrane protein (DRAP27) in monkey Vero cells. A cDNA encoding DRAP27 was isolated, and its nucleotide sequence was determined. The deduced amino acid sequence revealed that DRAP27 is the monkey homologue of human CD9 antigen. DRAP27 is recognized by CD9 antibodies. A human-mouse hybrid cell line (3279-10) possessing human chromosome 5, sensitive to DT, but not expressing CD9 antigen, was used for transfection experiments with DRAP27. When the cloned cDNA encoding DRAP27 was transiently expressed in 3279-10 cells, the total DT binding capacity was three to four times higher than that of untransfected controls. Transfectants stably expressing DRAP27 have an increased number of DT binding sites on the cell surface. Furthermore, the transfectants are 3-25 times more sensitive to DT than untransfected cells, and the sensitivity of these cells to DT is correlated with the number of DRAP27 molecules on the surface. However, when the cloned cDNA was introduced into mouse cell lines that do not express DT receptors, neither an increased DT binding nor enhancement of DT sensitivity was observed. Hence, we conclude that DRAP27 itself does not bind DT, but serves to increase DT binding and consequently enhances DT sensitivity of cells that have DT receptors. 12 proteins related to DRAP27/CD9 antigen were found through homology search analysis. These proteins appear to belong to a new family of transmembrane proteins.

scholarly journals Improved Specificity of a Multiplex Immunoassay for Quantitation of Anti-Diphtheria Toxin Antibodies with the Use of Diphtheria Toxoid

2011 ◽  
Vol 18 (7) ◽  
pp. 1183-1186 ◽  
Author(s):  
Pieter G. M. van Gageldonk ◽  
Christina von Hunolstein ◽  
Fiona R. M. van der Klis ◽  
Guy A. M. Berbers
Keyword(s):  
Diphtheria Toxin ◽  
Neutralization Test ◽  
Diphtheria Toxoid ◽  
Inhibition Assay ◽  
Nonspecific Binding ◽  
Serum Samples ◽  
Multiplex Immunoassay ◽  
Toxin Binding ◽  
Binding Inhibition ◽  
Binding Inhibition Assay

ABSTRACTA nonspecific binding of antibodies to diphtheria toxin, especially in adult serum samples, was observed in our diphtheria-tetanus-pertussis multiplex immunoassay (DTaP4 MIA). This can be significantly reduced by the use of diphtheria toxoid, achieving a good correlation with the Vero cell neutralization test and the toxin binding inhibition assay.

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