Pore-forming proteins (PFTs, also known as pore-forming toxins) are usually produced by bacteria, and include a number of protein exotoxins but may also be produced by other organisms such as apple snails that produce perivitellin-2 or earthworms, who produce lysenin. They are frequently cytotoxic (i.e., they kill cells), as they create unregulated pores in the membrane of targeted cells.
Types PFTs can be divided into two categories, depending on the alpha-helical or beta-barrel architecture of their transmembrane channel that can consist either of
Alpha-pore-forming toxins e.g., Haemolysin E family, actinoporins, Corynebacterial porin B, Cytolysin A of E. coli. Beta-barrel pore-forming toxins e.g. α-Hemolysin (Fig 1), PVL – Panton-Valentine leukocidin, various insecticidal toxins. Other categories:
Large beta-barrel pore-forming toxins MACPF and Cholesterol-dependent cytolysins (CDCs), gasdermin Binary toxins e.g., Anthrax toxin, Pleurotolysin Small pore-forming toxins e.g., Gramicidin A According to TCDB, there are following families of pore-forming toxins:
1.C.3 α-Hemolysin (αHL) family: 1.C.4 Aerolysin family 1.C.5 ε-Toxin family 1.C.11 RTX-toxin superfamily 1.C.12 Membrane attack complex/perforin superfamily 1.C.13 Leukocidin family 1.C.14 Cytohemolysin (CHL) family 1.C.39 Thiol-activated cholesterol-dependent cytolysin family 1.C.43 Lysenin family 1.C.56 Pseudomonas syringae HrpZ cation channel family 1.C.57 Clostridial cytotoxin family 1.C.74 Snake cytotoxin (SCT) family 1.C.97 Pleurotolysin pore-forming family
Beta-pore-forming toxins
β-PFTs are so-named because of their structural characteristics: they are composed mostly of β-strand-based domains. They have divergent sequences, and are classified by Pfam into a number of families including Leukocidins, Etx-Mtx2, Toxin-10, and aegerolysin. X-ray crystallographic structures have revealed some commonalities: α-hemolysin and Panton-Valentine leukocidin S are structurally related. Similarly, aerolysin and clostridial epsilon-toxin. and Mtx2 are linked in the Etx/Mtx2 family. The ß-PFTs include a number of toxins of commercial interest for the control of pest insects. These toxins are potent but also highly specific to a limited range of target insects, making them safe biological control agents. Insecticidal members of the Etx/Mtx2 family include Mtx2 and Mtx3 from Lysinibacillus sphaericus that can control mosquito vectors of human diseases and also Cry15, Cry23, Cry33, Cry38, Cry45, Cry51, Cry60, Cry64 and Cry74 from Bacillus thuringiensis that control a range of insect pests that can cause great losses to agriculture. Insecticidal toxins in the Toxin_10 family show an overall similarity to the aerolysin and Etx/Mtx2 toxin structures but differ in two notable features. While all of these toxins feature a head domain and a larger, extended beta-sheet tail domain, in the Toxin_10 family, the head is formed exclusively from the N-terminal region of the primary amino acid sequence whereas regions from throughout the protein sequence contribute to the head domain in Etx/Mtx2 toxins. In addition, the head domains of the Toxin_10 proteins show lectin-like features of carbohydrate binding domains. The only reported natural targets of Toxin_10 proteins are insects. With the exception of Cry36 and Cry78, the Toxin_10 toxins appear to act as two-part, binary toxins. The partner proteins in these combinations may belong to different structural groups, depending on the individual toxin: two Toxin_10 proteins (BinA and BinB) act together in the Bin mosquitocidal toxin of Lysinibacillus sphaericus; the Toxin_10 Cry49 is co-dependent on the 3-domain toxin family member Cry48 for its activity against Culex mosquito larvae; and the Bacillus thuringiensis Toxin_10 protein Cry35 interacts with the aegerolysin family Cry34 to kill Western Corn Rootworm. This toxin pair has been included in insect resistant plants such as SmartStax corn.
Mode of action
β-PFTs are dimorphic proteins that exist as soluble monomers and then assemble to form multimeric assemblies that constitute the pore. Figure 1 shows the pore-form of α-hemolysin, the first crystal structure of a β-PFT in its pore-form. 7 α-hemolysin monomers come together to create the mushroom-shaped pore. The 'cap' of the mushroom sits on the surface of the cell, and the 'stalk' of the mushroom penetrates the cell membrane, rendering it permeable (see later). The 'stalk' is composed of a 14-strand β-barrel, with two strands donated from each monomer. A structure of the Vibrio cholerae cytolysin PDB: 3O44 in the pore form is also heptameric; however, Staphylococcus aureus gamma-hemolysin PDB: 3B07 reveals an octomeric pore, consequently with a 16-strand 'stalk'. The Panton-Valentine leucocidin S structure PDB: 1T5R shows a highly related structure, but in its soluble monomeric state. This shows that the strands involved in forming the 'stalk' are in a very different conformation – shown in Fig 2. While the Bin toxin of Lysinibacillus sphaericus is able to form pores in artificial membranes and mosquito cells in culture, it also causes a series of other cellular changes including the uptake of toxin in recycling endosomes and the production of large, autophagic vesicles and the ultimate cause of cell death may be apoptotic. Similar effects on cell biology are also seen with other Toxin_10 activities but the roles of these events in toxicity remain to be established.
Assembly The transition between soluble monomer and membrane-associated protomer to oligomer is not a trivial one: It is believed that β-PFTs, follow as similar assembly pathway as the CDCs (see § Cholesterol-dependent cytolysins later), in that they must first assemble on the cell-surface (in a receptor-mediated fashion in some cases) in a pre-pore state. Following this, the large-scale conformational change occurs in which the membrane spanning section is formed and inserted into the membrane. The portion entering the membrane, referred to as the head, is usually apolar and hydrophobic, this produces an energetically favorable insertion of the pore-forming toxin.
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![Pore-forming toxin: a) The structure of perfringolysin O[31] and b) the structure of PluMACPF.[32] In both proteins, the two small clusters of α-helices that unwind and pierce the membrane are in pink. (PDB: 1PFO, 2QP2)](https://upload.wikimedia.org/wikipedia/commons/thumb/e/e7/PFO_and_Plu-MACPF.png/500px-PFO_and_Plu-MACPF.png?utm_source=en.wikipedia.org&utm_campaign=parser&utm_content=thumbnail)
