Expression of recombinant Clostridium difficile toxin A using the Bacillus megaterium system

Biochemical and Biophysical Research Communications

Volumn 307, Issue 3, 2003, Pages 584-588

  Burger, Silke ^a   Tatge, Helma ^a   Hofmann, Fred ^a   Genth, Harald ^a   Just, Ingo ^a   Gerhard, Ralf ^a  

^a HANNOVER MEDICAL SCHOOL   (Germany)

Author keywords

Cytotoxicity assay; Endotoxin free expression system; Glucosyltransferase; pWH1520 vector

Indexed keywords

CLOSTRIDIUM DIFFICILE TOXIN A; GLUCOSYLTRANSFERASE; HISTIDINE; NICKEL; RECOMBINANT ENZYME; THYROGLOBULIN; TOXIN ANTIBODY;

AFFINITY CHROMATOGRAPHY; AMINO ACID SEQUENCE; ARTICLE; BACILLUS MEGATERIUM; BACTERIAL GENETICS; BACTERIAL STRAIN; CARBOXY TERMINAL SEQUENCE; CLONING VECTOR; CLOSTRIDIUM DIFFICILE; CONTROLLED STUDY; CYTOTOXICITY; ENZYME ACTIVITY; ENZYME ANALYSIS; ENZYME STRUCTURE; GENE CONSTRUCT; GENE EXPRESSION SYSTEM; GENE SEQUENCE; HUMAN; HUMAN CELL; IMMUNOASSAY; IN VITRO STUDY; MOLECULAR CLONING; NONHUMAN; PRIORITY JOURNAL; PROTEIN EXPRESSION; PROTEIN FUNCTION; PROTEIN PURIFICATION; SEQUENCE ANALYSIS;

BACILLUS MEGATERIUM; CLOSTRIDIUM; CLOSTRIDIUM DIFFICILE; UNIDENTIFIED CLONING VECTOR; VECTORS;

EID: 0041670743     PISSN: 0006291X     EISSN: None     Source Type: Journal    
DOI: 10.1016/S0006-291X(03)01234-8     Document Type: Article

References (19)

1. Mylonakis E., Ryan E.T., Calderwood S.B. Clostridium difficile-associated diarrhea. Arch. Int. Med. 161:2001;525-533.
2. Kelly C.P., Pothoulakis C., LaMont J.T. Clostridium difficile colitis. New Engl. J. Med. 330:1994;257-262.
3. Chaves-Olarte E., Weidmann M., Von Eichel-Streiber C., Thelestam M. Toxins A and B from Clostridium difficile differ with respect to enzymatic potencies, cellular substrate specificities, and surface binding to cultured cells. J. Clin. Invest. 100:1997;1734-1741.
4. Just I., Wilm M., Selzer J., Rex G., Von Eichel-Streiber C., Mann M., Aktories K. The enterotoxin from Clostridium difficile (ToxA) monoglucosylates the Rho proteins. J. Biol. Chem. 270:1995;13932-13936.
5. Krivan H.C., Wilkins T.D. Purification of Clostridium difficile toxin A by affinity chromatography on immobilized thyroglobulin. Infect. Immun. 55:1987;1873-1877.
6. Roberts A.K., Shone C.C. Modification of surface histidine residues abolishes the cytotoxic activity of Clostridium difficile toxin A. Toxicon. 39:2001;325-333.
7. Sauerborn M., Leukel P., Von Eichel-Streiber C. The C-terminal ligand-binding domain of Clostridium difficile toxin A (TcdA) abrogates TcdA-specific binding to cells and prevents mouse lethality. FEMS Microbiol. Lett. 155:1997;45-54.
8. Frisch C., Gerhard R., Aktories K., Hofmann F., Just I. The complete receptor-binding domain of Clostridium difficile toxin A is required for endocytosis. Biochem. Biophys. Res. Commun. 300:2003;706-711.
9. Makoff A.J., Ballantine S.P., Smallwood A.E., Neil F. Expression of tetanus toxin fragment C in E. coli: its purification and potential use as a vaccine. Biotechnology. 7:1989;1043-1046.
10. Tang-Feldman Y.J., Ackermann G., Henderson J.P., Silva J., Cohen S.H. One-step cloning and expression of Clostridium difficile toxin B gene (tcdB). Mol. Cell Probes. 16(3):2002;179-183.
11. Vary P.S. Prime time for Bacillus megaterium. Microbiology. 140:1994;1001-1013.
12. Just I., Selzer J., Hofmann F., Aktories K. Clostridium difficile toxin B as a probe for Rho GTPases. Aktories K. Bacterial Toxins - Tools in Cell Biology and Pharmacology. 1997;159-168 Chapman & Hall, Weinheim.
13. Genth H., Selzer J., Busch C., Dumbach J., Hofmann F., Aktories K., Just I. New method to generate enzymatically deficient Clostridium difficile toxin B as an antigen for immunization. Infect. Immun. 68:2000;1094-1101.
14. Genth H., Aktories K., Just I. Monoglucosylation of RhoA at Threonine-37 blocks cytosol-membrane cycling. J. Biol. Chem. 274:1999;29050-29056.
15. Kyriakis J.M., Avruch J. Mammalian mitogen-activated protein kinase signal transduction pathways activated by stress and inflammation. Physiol. Rev. 81(2):2003;807-869.
16. Ono K., Han J. The p38 signal transduction pathway: activation and function. Cell. Signal. 12:2000;1-13.
17. Leong K.G., Karsan A. Signaling pathways mediated by tumor necrosis factor alpha. Histol. Histopathol. 15(4):2000;1303-1325.
18. Woo C.H., Kim J.H. Rac GTPase activity is essential for lipopolysaccharide signaling to extracellular signal-regulated kinase and p38 MAP kinase activation in rat-2 fibroblasts. Mol. Cells. 13(3):2002;470-475.
19. Sanlioglu S., Williams C.M., Samavati L., Butler N.S., Wang G., McCray P.B., Ritchi T.C. Jr., Hunninghake G.W., Zandi E., Engelhardt J.F. Lipopolysaccharide induces Rac1-dependent reactive oxygen species formation and coordinates tumor necrosis factor-alpha secretion through IKK regulation of NF-κB. J. Biol. Chem. 276(32):2001;30188-30198.