1.C.7 The Diphtheria Toxin (DT) Family The DT family consists of a single protein, diphtheria toxin (DT). DT is synthesized as a 535 amino acyl residue protein encoded by the tox gene of corynebacteriophage beta in Corynebacterium diphtheriae. The protein is secreted as a single polypeptide chain in a leader peptide-dependent process and is then cleaved between arg-193 and ser-194 by extracellular trypsin-like proteases. This cleavage event yields the N-terminal A chain that contains the NAD⁺:diphtheramide ADP ribosyl transferase (EC 2.4.2.36) activity, and the C-terminal B chain that forms the transmembrane channel that allows the A chain to enter the cytoplasm of the eukaryotic cell where the latter exerts its toxic effect by ADP ribosylating a histidyl residue on the N-terminal region of elongation factor 2 (EF2). In the extracellular environment, the A and B chains are held together by a disulfide bridge. Diphtheria toxin serves as the prototype for two chain (or multidomain) bacterial and plant toxins. The B chain binds to the external plasma membrane receptor as a prelude to channel formation. The C-terminal half of the B chain is the receptor binding domain while the N-terminal half as well as part of the binding domain comprise the channel. While the translocation process and the structure of the membrane embedded channel are not understood in detail, translocation has been reported to require energy. Lai et al. (2008) reported that of the deeply inserted helices in the Diphtheria toxin T domain: helices 5, 8, and 9, interact strongly and promote pore formation, while helices 6/7 limit pore formation. Other well-characterized toxins that have an AB structure, where B translocates A, include botulinum neurotoxins A-G, several chlostridial neurotoxins, two anthrax neurotoxins, and tetanus neurotoxin. Membrane insertion of the B chain and channel formation often occur after phagocytosis in vesicles or the endoplasmic reticulum where acid pH induces the conformational change that accompanies insertion into the endosomal membrane. After disulfide reduction, exposure to the neutral pH of the cytosol may trigger the final translocation of the A-chain. DT appears to form oligomers with variable stoichiometry and pore size. The greater the number of monomers, the larger the pore size (Sharpe and London, 1999). The diphtheria toxin T domain helps translocate the A chain of the toxin across membranes. The membrane topography of the diphtheria toxin T domain linked to the A chain revealed a transient transmembrane hairpin and potential translocation mechanisms (Wang and London, 2009). The transport reaction catalyzed by DT is: A-chain (out) → A-chain (in).
References:
Blanke, S.R. (2006). Portals and pathways: principles of bacterial toxin entry into host cells. Microbe 1: 26-31.
D’Silva, P.R. and A.K. Lala (2000). Organization of diphtheria toxin in membranes, a hydrophobic photolabeling study. J. Biol. Chem. 275: 11771-11777.
Greenfield, L., M.J. Bjorn, G. Horn, D. Fong, G.A. Buck, R.J. Collier and D.A. Kaplan (1983). Nucleotide sequence of the structural gene for diphtheria toxin carried by corynebacteriophage beta. Proc. Natl. Acad. Sci. USA 80: 6853-6857.
Kachel, K., J. Ren, R.J. Collier and E. London (1998). Identifying transmembrane states and defining the membrane insertion boundaries of hydrophobic helices in membrane-inserted diphtheria toxin T domain. J. Biol. Chem. 273: 22950-22956.
Lai, B., G. Zhao, and E. London. (2008). Behavior of the deeply inserted helices in diphtheria toxin T domain: helices 5, 8, and 9 interact strongly and promote pore formation, while helices 6/7 limit pore formation. Biochemistry 47: 4565-4574.
Lesieur, C., B. Vécsey-Semjén, L. Abrami, M. Fivaz and F. Gisou van der Goot (1997). Membrane insertion: the strategies of toxins. Mol. Membr. Biol. 14: 45-64.
London, E. (1992). Diphtheria toxin: membrane interaction and membrane translocation. Biochim. Biophys. Acta 1113: 25-51.
Misler, S. (1983). Gating of ion channels made by a diphtheria toxin fragment in phospholipid bilayer membranes. Proc. Natl. Acad. Sci. USA 80: 4320-4324.
Montecucco, C. (1998). Protein toxins and membrane transport. Curr. Opin. Cell Biol. 10: 530-536.
Neville, Jr., D.M. and T.H. Hudson (1986). Transmembrane transport of diphtheria toxin, related toxins and colicins. Ann. Rev. Biochem. 55: 195-224.
Sharpe, J.C. and E. London (1999). Diphtheria toxin forms pores of different sizes depending on its concentration in membranes: probable relationship to oligomerization. J. Memb. Biol. 171: 209-221.
Wang, J. and E. London. (2009). The membrane topography of the diphtheria toxin T domain linked to the a chain reveals a transient transmembrane hairpin and potential translocation mechanisms. Biochemistry 48: 10446-10456.
Examples:
TC#	Name	Organismal Type	Example
1.C.7.1.1	Diphtheria toxin (DT)	Corynebacterium diphtheriae phage beta	DT of corynebacteriophage beta