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Gas phase characterization of the noncovalent quaternary structure of Cholera toxin and the Cholera toxin B subunit pentamer

By Jonathan P. Williams, Daniel C. Smith, Brian N. Green, Brian D. Marsden, Keith R. Jennings, L. M. (Lynne M.) Roberts and James H. Scrivens

Abstract

Cholera toxin (CTx) is an AB5 cytotonic protein that has medical relevance in cholera and as a novel mucosal adjuvant. Here, we report an analysis of the noncovalent homopentameric complex of CTx B chain (CTx B5) using electrospray ionization triple quadrupole mass spectrometry and tandem mass spectrometry and the analysis of the noncovalent hexameric holotoxin usingelectrospray ionization time-of-flight mass spectrometry over a range of pH values that correlate with those encountered by this toxin after cellular uptake. We show that noncovalent interactions within the toxin assemblies were maintained under both acidic and neutral conditions in the gas phase. However, unlike the related Escherichia coli Shiga-like toxin B5 pentamer (SLTx B), the CTx B5 pentamer was stable at low pH, indicating that additional interactions must be present within the latter. Structural comparison of the CTx B monomer interface reveals an additional α-helix that is absent in the SLTx B monomer. In silico energy calculations support interactions between this helix and the adjacent monomer. These data provide insight into the apparent stabilization of CTx B relative to SLTx B

Topics: QR180
Publisher: Biophysical Society
Year: 2006
OAI identifier: oai:wrap.warwick.ac.uk:924

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  1. 4 A ˚ crystal structure of cholera toxin B subunit pentamer: choleragenoid. doi
  2. (2003). A class of mutant CHO cells resistant to cholera toxin rapidly degrades the catalytic polypeptide of cholera toxin and exhibits increased endoplasmic reticulum-associated degradation. doi
  3. (1995). Analysis of Shiga toxin subunit association by using hybrid A polypeptides and site-specific mutagenesis.
  4. (1989). Binding of Vibrio cholera toxin and the heat-labile enterotoxin of Escherichia coli to GM1, derivatives of GM1, and nonlipid oligosaccharide polyvalent ligands. doi
  5. (1999). Characterisation of cholera toxin by liquid chromatography—electrospray mass spectrometry. doi
  6. (1986). Characterization of the receptor for cholera toxin and Escherichia coli heatlabile toxin in rabbit intestinal brush borders.
  7. (2005). Cholera and Shiga toxin B-subunits: thermodynamic and structural considerations for function and biomedical applications. doi
  8. (1988). Cholera toxin and pertussis toxin stimulate prostaglandin E2 synthesis in a murine macrophage cell line.
  9. (2004). Cholera toxin toxicity does not require functional Arf6- and dynamin-dependent endocytic pathways. doi
  10. (2004). Cholera toxin: a paradigm for multi-functional engagement of cellular mechanisms (Review). doi
  11. (1974). Cholera toxin: interaction of subunits with ganglioside GM1. doi
  12. (1993). Comparison of the B-pentamers of heat-labile enterotoxin and verotoxin-1: two structures with remarkable similarity and dissimilarity. doi
  13. (1997). Conformational changes in cholera toxin B subunit-ganglioside GM1 complexes are elicited by environmental pH and evoke changes in membrane structure. doi
  14. (1982). Conformational changes in subunit A of cholera toxin following the binding of ganglioside to subunit doi
  15. (1993). Convention on the prohibition of the development, production, stockpiling and use of chemical weapons and on their destruction. doi
  16. Crystal structure of the cell-binding B oligomer of verotoxin1 from E.
  17. (1994). Crystal structure of the holotoxin from Shigella dysenteriae at 2.5 A ˚ resolution. doi
  18. (1972). Effect of cholera enterotoxin on ion transport across isolated ileal mucosa. doi
  19. (1970). Elevated concentration of adenosine 39:59-cyclic monophosphate in intestinal mucosa after treatment with cholera toxin. doi
  20. (1993). Epidemiology of diarrhoeal disease: implications for control by vaccines. doi
  21. (1985). Facile identification of protein sequences by mass spectrometry. B subunit of Vibrio cholerae classical biotype Inaba 569B toxin. doi
  22. (1995). Fluorescence analysis of the interaction between ganglioside GM1-containing phospholipid vesicles and the B subunit of cholera toxin. doi
  23. (2003). Gangliosides that associate with lipid rafts mediate transport of cholera and related toxins from the plasma membrane to endoplasmic reticulum. doi
  24. (1978). Genetic evidence that cholera toxin substrates are regulatory components of adenylate cyclase. doi
  25. (1994). ICM—a new method for protein modeling and design: applications to docking and structure prediction from the distorted native conformation. doi
  26. (1972). Intestinal adenylcyclase activity in canine cholera: correlation with fluid accumulation. doi
  27. (1989). Intestinal electrolyte transport and diarrheal disease (2). doi
  28. (2005). New drug targets for cholera therapy. Trends Pharmacol. doi
  29. (2005). Noncovalent Shiga-like toxin assemblies: characterization by means of mass spectrometry and tandem mass spectrometry. doi
  30. (2004). Pathways followed by protein toxins into cells. doi
  31. (1991). Phosphorylation of the R domain by cAMP-dependent protein kinase regulates the CFTR chloride channel. doi
  32. (2001). Protein disulfide isomerase acts as a redox-dependent chaperone to unfold cholera toxin. doi
  33. (2002). Retro-translocation of proteins from the endoplasmic reticulum into the cytosol. doi
  34. (2004). Retrograde transport of cholera toxin from the plasma membrane to the endoplasmic reticulum requires the transGolgi network but not the Golgi apparatus in Exo2-treated cells. doi
  35. (2004). Retrograde transport of cholera toxin into the ER of host cells. doi
  36. (1971). Stimulation of intestinal adenyl cyclase by cholera toxin. doi
  37. (1990). Structure, stability, and receptor interaction of cholera toxin as studied by Fourier-transform infrared spectroscopy. doi
  38. (1998). The 1.25 A ˚ resolution refinement of the cholera toxin B-pentamer: evidence of peptide backbone strain at the receptorbinding site. doi
  39. (2003). The intracellular voyage of cholera toxin: going retro. Trends Biochem. doi
  40. (2001). The observation of multivalent complexes of Shiga-like toxin with globotriaoside and the determination of their stoichiometry by nanoelectrospray Fourier-transform ion cyclotron resonance mass spectrometry. doi
  41. (1971). The possible role of cyclic AMP in some actions of cholera toxin. doi
  42. (2000). The protein data bank. Nucleic Acids Res. doi
  43. (1995). The three-dimensional crystal structure of cholera toxin. doi
  44. (2001). Thermal decomposition of a gaseous multiprotein complex studied by blackbody infrared radiative dissociation. Investigating the origin of the asymmetric dissociation behavior. doi
  45. (1988). Thermal stability and intersubunit interactions of cholera toxin in solution and in association with its cellsurface receptor ganglioside GM1. doi
  46. (2003). Thermodynamic analysis of the structural stability of the Shiga toxin B-subunit. doi
  47. (2002). Unfolded cholera toxin is transferred to the ER membrane and released from protein disulfide isomerase upon oxidation by Ero1. doi

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