Diphtheria toxin is an exotoxin secreted by mainly by Corynebacterium diphtheriae but also by Corynebacterium ulcerans and Corynebacterium pseudotuberculosis. the pathogenic bacterium that causes diphtheria. The toxin gene is encoded by a prophage called corynephage β. The toxin causes the disease in humans by gaining entry into the cell cytoplasm and inhibiting protein synthesis. Structure Diphtheria toxin is a single polypeptide chain of 535 amino acids consisting of two subunits linked by disulfide bridges, known as an A-B toxin. Binding to the cell surface of the B subunit (the less stable of the two subunits) allows the A subunit (the more stable part of the protein) to penetrate the host cell. The crystal structure of the diphtheria toxin homodimer has been determined to 2.5 Ångstrom resolution. The structure reveals a Y-shaped molecule consisting of three domains. Fragment A contains the catalytic C domain, and fragment B consists of the T and R domains: The amino-terminal catalytic domain, known as the C domain, has an unusual beta+alpha fold. The C domain blocks protein synthesis by transfer of ADP-ribose from NAD to a diphthamide residue of eukaryotic elongation factor 2 (eEF-2). A central translocation domain, known as the T domain or TM domain, has a multi-helical globin-like fold with two additional helices at the amino terminus but no counterpart to the first globin helix. This domain is thought to unfold in the membrane. A pH-induced conformational change in the T domain triggers insertion into the endosomal membrane and facilitates the transfer of the C domain into the cytoplasm. A carboxy-terminal receptor-binding domain, known as the R domain, has a beta-sandwich fold consisting of nine strands in two sheets with Greek-key topology; it is a subclass of immunoglobulin-like fold. The R domain binds to a cell surface receptor, permitting the toxin to enter the cell by receptor-mediated endocytosis. Mechanism Processing The leader region is cleaved during secretion. Proteolytic nicking separates A and B subunits, which remain joined by disulfide bonds until they reach the cytosol. The toxin binds to heparin-binding epidermal growth factor precursor (HB-EGF). The complex undergoes endocytosis by the host cell. Acidification inside the endosome induces translocation of the A subunit into the cytosol. Disulfide bonds are broken. The B subunit remains in the endosome as a pore. The A subunit ADP-ribosylates host eEF-2, which is required for protein synthesis; when it is inactivated, the host cannot make protein and thus