DF/HCC Program Affiliation

Cancer Cell Biology

Research Abstract

The symptoms of many bacterial diseases are due largely to the actions of toxic proteins released by the bacteria (diphtheria, anthrax, cholera, and tetanus toxins are well known examples). We study these toxins with the goals of understanding the biochemical basis of bacterial disease; gaining insight into how proteins cross membranes; and developing new pharmaceuticals. The most potent toxins act by penetrating into mammalian cells and modifying target substrates within the cytosol. Their actions generally involve four steps: (i) binding to receptors; (ii) endocytosis of the toxin-receptor complex; (iii) translocation of the enzymic part across a membrane, into the cytosol; and (iv) enzymic methods to generate detailed models of each step in toxin action. How these structurally unrelated toxins insert into membranes under the influence of the low pH of the endosomal compartment, and translocate their enzymic moieties across the endosomal membrane represents a problem of broad interest. Crystallographic structures of the native proteins provide a framework for our studies. Genetically modified, nontoxic forms of these toxins may be used as transporters for heterologous proteins and peptides into cells. We are using anthrax toxin to engineer a new type of vaccines that stimulates the formation of cytotoxic T lymphocytic (CTL) responses, as opposed to antibody formation. CTL-based vaccines may be useful against a variety of diseases which have heretofore been thought to be beyond the realm of vaccination.

Publications

• Lacy DB, Lin HC, Melnyk RA, Schueler-Furman O, Reither L, Cunningham K, Baker D, Collier RJ.A model of anthrax toxin lethal factor bound to protective antigen.Proc Natl Acad Sci U S A 2005 Nov 8;102(45):16409-14.
  16251269
• Wolfe JT, Krantz BA, Rainey GJ, Young JA, Collier RJ.Whole-cell voltage clamp measurements of anthrax toxin pore current.J Biol Chem 2005 Sep 23.
  16183642
• Krantz BA, Melnyk RA, Zhang S, Juris SJ, Lacy DB, Wu Z, Finkelstein A, Collier RJ.A phenylalanine clamp catalyzes protein translocation through the anthrax toxin pore.Science 2005 Jul 29;309(5735):777-81.
  16051798
• Zhang S, Finkelstein A, Collier RJ.Evidence that translocation of anthrax toxin's lethal factor is initiated by entry of its N terminus into the protective antigen channel.Proc Natl Acad Sci U S A 2004 Nov 30;101(48):16756-61.
  15548616
• Kim C, Wilcox-Adelman S, Sano Y, Tang WJ, Collier RJ, Park JM.Antiinflammatory cAMP signaling and cell migration genes co-opted by the anthrax bacillus.Proc Natl Acad Sci U S A 2008 Apr 22;105(16):6150-5.
  18427110
• Sun J, Lang AE, Aktories K, Collier RJ.Phenylalanine-427 of anthrax protective antigen functions in both pore formation and protein translocation.Proc Natl Acad Sci U S A 2008 Mar 18;105(11):4346-51.
  18334631
• Bolcome RE 3rd, Sullivan SE, Zeller R, Barker AP, Collier RJ, Chan J.Anthrax lethal toxin induces cell death-independent permeability in zebrafish vasculature.Proc Natl Acad Sci U S A 2008 Feb 19;105(7):2439-44.
  18268319
• Wimalasena DS, Cramer JC, Janowiak BE, Juris SJ, Melnyk RA, Anderson DE, Kirk KL, Collier RJ, Bann JG.Effect of 2-fluorohistidine labeling of the anthrax protective antigen on stability, pore formation, and translocation.Biochemistry 2007 Dec 25;46(51):14
  18044973
• Rainey GJ, Wigelsworth DJ, Ryan PL, Scobie HM, Collier RJ, Young JA.Receptor-specific requirements for anthrax toxin delivery into cells.Proc Natl Acad Sci U S A 2005 Sep 13;102(37):13278-83.
  16141341
• Scobie HM, Thomas D, Marlett JM, Destito G, Wigelsworth DJ, Collier RJ, Young JA, Manchester M.A soluble receptor decoy protects rats against anthrax lethal toxin challenge.J Infect Dis 2005 Sep 15;192(6):1047-51.
  16107958
• Qa'dan M, Christensen KA, Zhang L, Roberts TM, Collier RJ.Membrane insertion by anthrax protective antigen in cultured cells.Mol Cell Biol 2005 Jul;25(13):5492-8.
  15964805
• Christensen KA, Krantz BA, Melnyk RA, Collier RJ.Interaction of the 20 kDa and 63 kDa fragments of anthrax protective antigen: kinetics and thermodynamics.Biochemistry 2005 Jan 25;44(3):1047-53.
  15654761
• Krantz BA, Trivedi AD, Cunningham K, Christensen KA, Collier RJ.Acid-induced unfolding of the amino-terminal domains of the lethal and edema factors of anthrax toxin.J Mol Biol 2004 Nov 26;344(3):739-56.
  15533442
• Boll W, Ehrlich M, Collier RJ, Kirchhausen T.Effects of dynamin inactivation on pathways of anthrax toxin uptake.Eur J Cell Biol 2004 Jul;83(6):281-8.
  15511085
• Zhang S, Udho E, Wu Z, Collier RJ, Finkelstein A.Protein translocation through anthrax toxin channels formed in planar lipid bilayers.Biophys J 2004 Dec;87(6):3842-9.
  15377524
• Lacy DB, Wigelsworth DJ, Melnyk RA, Harrison SC, Collier RJ.Structure of heptameric protective antigen bound to an anthrax toxin receptor: a role for receptor in pH-dependent pore formation.Proc Natl Acad Sci U S A 2004 Sep 7;101(36):13147-51.
  15326297
• Pimental RA, Christensen KA, Krantz BA, Collier RJ.Anthrax toxin complexes: heptameric protective antigen can bind lethal factor and edema factor simultaneously.Biochem Biophys Res Commun 2004 Sep 10;322(1):258-62.
  15313199
• Ladokhin AS, Legmann R, Collier RJ, White SH.Reversible refolding of the diphtheria toxin T-domain on lipid membranes.Biochemistry 2004 Jun 15;43(23):7451-8.
  15182188
• Zhang S, Cunningham K, Collier RJ.Anthrax protective antigen: efficiency of translocation is independent of the number of ligands bound to the prepore.Biochemistry 2004 May 25;43(20):6339-43.
  15147218
• Mourez M, Collier RJ.Use of phage display and polyvalency to design inhibitors of protein-protein interactions.Methods Mol Biol 2004;261:213-28.
  15064461