scholarly journals KDEL Receptor (Erd2p)-mediated Retrograde Transport of the Cholera Toxin A Subunit from the Golgi Involves COPI, p23, and the COOH Terminus of Erd2p

1998 ◽  
Vol 143 (3) ◽  
pp. 601-612 ◽  
Author(s):  
Irina Majoul ◽  
Kai Sohn ◽  
Felix Theodor Wieland ◽  
Rainer Pepperkok ◽  
Mariagrazia Pizza ◽  
...  
Keyword(s):  
Coat Protein ◽  
Cholera Toxin ◽  
Retrograde Transport ◽  
Vero Cells ◽  
Toxin A ◽  
P24 Protein ◽  
A Subunit ◽  
Intermediate Compartment ◽  
Camp Levels ◽  
Kdel Receptor

A cholera toxin mutant (CTX–K63) unable to raise cAMP levels was used to study in Vero cells the retrograde transport of the toxin A subunit (CTX-A–K63), which possesses a COOH-terminal KDEL retrieval signal. Microinjected GTP-γ-S inhibits the internalization as well as Golgi–ER transport of CTX-A–K63. The appearance of CTX-A–K63 in the Golgi induces a marked dispersion of Erd2p and p53 but not of the Golgi marker giantin. Erd2p is translocated under these conditions most likely to the intermediate compartment as indicated by an increased colocalization of Erd2p with mSEC13, a member of the mammalian coat protein II complex. IgGs as well as Fab fragments directed against Erd2p, β-COP, or p23, a new member of the p24 protein family, inhibit or block retrograde transport of CTX-A–K63 from the Golgi without affecting its internalization or its transport to the Golgi. Anti-Erd2p antibodies do not affect the binding of CTX-A to Erd2p, but inhibit the CTX-K63–induced translocation of Erd2p and p53.

scholarly journals Transport of an external Lys-Asp-Glu-Leu (KDEL) protein from the plasma membrane to the endoplasmic reticulum: studies with cholera toxin in Vero cells.

1996 ◽  
Vol 133 (4) ◽  
pp. 777-789 ◽  
Author(s):  
I V Majoul ◽  
P I Bastiaens ◽  
H D Söling
Keyword(s):  
Endoplasmic Reticulum ◽  
Cholera Toxin ◽  
Retrograde Transport ◽  
Vero Cells ◽  
Subcellular Fractionation ◽  
Pulse Treatment ◽  
Resistant Form ◽  
Immunofluorescence Studies ◽  
Intermediate Compartment ◽  
Kdel Sequence

The A2 chain of cholera toxin (CTX) contains a COOH-terminal Lys-Asp-Glu-Leu (KDEL) sequence. We have, therefore, analyzed by immunofluorescence and by subcellular fractionation in Vero cells whether CTX can used to demonstrate a retrograde transport of KDEL proteins from the Golgi to the ER. Immunofluorescence studies reveal that after a pulse treatment with CTX, the CTX-A and B subunits (CTX-A and CTX-B) reach Golgi-like structures after 15-20 min (maximum after 30 min). Between 30 and 90 min, CTX-A (but not CTX-B) appear in the intermediate compartment and in the ER, whereas the CTX-B are translocated to the lysosomes. Subcellular fractionation studies confirm these results: after CTX uptake for 15 min, CTX-A is associated only with endosomal and Golgi compartments. After 30 min, a small amount of CTX-A appears in the ER in a trypsin-resistant form, and after 60 min, a significant amount appears. CTX-A seems to be transported mainly in its oxidized form (CTX-A1-S-S-CTX-A2) from the Golgi to the ER, where it becomes slowly reduced to form free CTX A1 and CTX-A2, as indicated by experiments in which cells were homogenized 30 and 90 min after the onset of CTX uptake in the presence of N-ethylmaleimide. Nocodazol applied after accumulation of CTX in Golgi inhibits the appearance of CTX-A in the ER and delays the increase of 3',5'cAMP, indicating the participation of microtubules in the retrograde Golgi-ER transport.

scholarly journals Expression and mutagenesis of recombinant cholera toxin A subunit

1994 ◽  
Vol 17 (5) ◽  
pp. 339-346 ◽  
Author(s):  
Kirsten L. Vadheim ◽  
Yogendra Singh ◽  
Jerry M. Keith
Keyword(s):  
Cholera Toxin ◽  
Toxin A ◽  
A Subunit

scholarly journals p97 Is in a Complex with Cholera Toxin and Influences the Transport of Cholera Toxin and Related Toxins to the Cytoplasm

2005 ◽  
Vol 280 (16) ◽  
pp. 15865-15871 ◽  
Author(s):  
Ramzey J. AbuJarour ◽  
Seema Dalal ◽  
Phyllis I. Hanson ◽  
Rockford K. Draper
Keyword(s):  
Endoplasmic Reticulum ◽  
Cholera Toxin ◽  
Vero Cells ◽  
Degradation Pathway ◽  
Endoplasmic Reticulum Membrane ◽  
Toxin A ◽  
Aaa Atpase ◽  
Endoplasmic Reticulum Associated Degradation ◽  
Protein Toxins ◽  
A Chain

Certain protein toxins, including cholera toxin, ricin, andPseudomonas aeruginosaexotoxin A, are transported to the lumen of the endoplasmic reticulum where they retro-translocate across the endoplasmic reticulum membrane to enter the cytoplasm. The mechanism of retrotranslocation is poorly understood but may involve the endoplasmic reticulum-associated degradation pathway. The AAA ATPase p97 (also called valosin-containing protein) participates in the retro-translocation of cellular endoplasmic reticulum-associated degradation substrates and is therefore a candidate to participate in the retrotranslocation of protein toxins. To investigate whether p97 functions in toxin delivery to the cytoplasm, we measured the sensitivity to toxins of cells expressing either wild-type p97 or a dominant ATPase-defective p97 mutant under control of a tetracycline-inducible promoter. The rate at which cholera toxin and related toxins entered the cytoplasm was reduced in cells expressing the ATPase-defective p97, suggesting that the toxins might interact with p97. To detect interaction, the cholera toxin A chain was immunoprecipitated from cholera toxin-treated Vero cells, and co-immunoprecipitation of p97 was assessed by immunoblotting. The immunoprecipitates contained both cholera toxin A chain and p97, evidence that the two proteins are in a complex. Altogether, these results provide functional and structural evidence that p97 participates in the transport of cholera toxin to the cytoplasm.

Anthrax toxin-mediated delivery of cholera toxin-A subunit into the cytosol of mammalian cells

2000 ◽  
Vol 32 (1) ◽  
pp. 69 ◽  
Author(s):  
Manju Sharma ◽  
Hemant Khanna ◽  
Naveen Arora ◽  
Yogendra Singh
Keyword(s):  
Cholera Toxin ◽  
Mammalian Cells ◽  
Anthrax Toxin ◽  
Toxin A ◽  
A Subunit

scholarly journals Sleep-inducing effect of substance P-cholera toxin A subunit in mice

2017 ◽  
Vol 659 ◽  
pp. 44-47 ◽  
Author(s):  
Mark R. Zielinski ◽  
Dmitry Gerashchenko
Keyword(s):  
Substance P ◽  
Cholera Toxin ◽  
Toxin A ◽  
A Subunit

scholarly journals A molecular specificity code for the three mammalian KDEL receptors

2007 ◽  
Vol 179 (6) ◽  
pp. 1193-1204 ◽  
Author(s):  
Irina Raykhel ◽  
Heli Alanen ◽  
Kirsi Salo ◽  
Jaana Jurvansuu ◽  
Van Dat Nguyen ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Coat Protein ◽  
Bimolecular Fluorescence Complementation ◽  
Reporter Construct ◽  
Human Proteins ◽  
Molecular Specificity ◽  
Intermediate Compartment ◽  
Kdel Receptor ◽  
Dependent Pathway ◽  
Construct System

AC-terminal KDEL-like motif prevents secretion of soluble endoplasmic reticulum (ER)–resident proteins. This motif interacts with KDEL receptors localized in the intermediate compartment and Golgi apparatus. Such binding triggers retrieval back to the ER via a coat protein I–dependent pathway. To date, two human KDEL receptors have been reported. Here, we report the Golgi localization of a third human KDEL receptor. Using a reporter construct system from a screen of 152 variants, we identified 35 KDEL-like variants that result in efficient ER localization but do not match the current Prosite motif for ER localization ([KRHQSA]-[DENQ]-E-L). We cloned 16 human proteins with one of these motifs and all were found in the ER. A subsequent screen by bimolecular fluorescence complementation determined the specificities of the three human KDEL receptors. Each KDEL receptor has a unique pattern of motifs with which it interacts. This suggests a specificity in the retrieval of human proteins that contain different KDEL variants.

scholarly journals Engineering of cholera toxin A-subunit for carriage of epitopes at its amino end

FEBS Letters ◽  
1997 ◽  
Vol 401 (1) ◽  
pp. 95-97 ◽  
Author(s):  
J Sanchez ◽  
R Argotte ◽  
A Buelna
Keyword(s):  
Cholera Toxin ◽  
Toxin A ◽  
A Subunit

scholarly journals Nasally administered cholera toxin A-subunit acts as a mucosal adjuvant

2003 ◽  
Vol 45 (1) ◽  
pp. 25-31 ◽  
Author(s):  
Emilio A. Campos ◽  
Jun Namikoshi ◽  
Satomi Maeba ◽  
Masafumi Yamamoto ◽  
Masahiko Fukumoto ◽  
...  
Keyword(s):  
Cholera Toxin ◽  
Toxin A ◽  
Mucosal Adjuvant ◽  
A Subunit
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