The Membrane Attack Complex-Perforin (MACPF) family is ubiquitously found in all kingdoms. They have diverse cellular roles, however MACPFs with pore-forming toxic function in venoms and poisons are very rare in animals. Here we present the structure of PmPV2, a MACPF toxin from the poisonous apple snail eggs, that can affect the digestive and nervous systems of potential predators. We report the three-dimensional structure of PmPV2, at 17.2 Å resolution determined by negative-stain electron microscopy and its solution structure by small angle X-ray scattering (SAXS). We found that PV2s differ from nearly all MACPFs in two respects: it is a dimer in solution and protomers combine two immune proteins into an AB toxin. The MACPF chain is linked by a single disulfide bond to a tachylectin chain, and two heterodimers are arranged head-to-tail by non-covalent forces in the native protein. MACPF domain is fused with a putative new Ct-accessory domain exclusive to invertebrates. The tachylectin is a six-bladed β-propeller, similar to animal tectonins. We experimentally validated the predicted functions of both subunits and demonstrated for the first time that PV2s are true pore-forming toxins. The tachylectin “B” delivery subunit would bind to target membranes, and then the MACPF “A” toxic subunit would disrupt lipid bilayers forming large pores altering the plasma membrane conductance. These results indicate that PV2s toxicity evolved by linking two immune proteins where their combined preexisting functions gave rise to a new toxic entity with a novel role in defense against predation. This structure is an unparalleled example of protein exaptation.
Bibliographical noteFunding Information:
This work was supported by funding from Ministry of Science and Technology of Argentina Grant (Agencia Nacional de Promoción Científica y Tecnológica, PICT 2014-0850 to HH and PICT 2013-0122 to SI.), Consejo Nacional de Investigaciones Científicas y Técnicas, CONICET (PIP 0051 to HH), General Research Fund of Hong Kong (HKBU 12,301,415 to JW Q), and by partial financial support from LNLS–Brazilian Synchrotron Light Laboratory/MCT (Project SAXS1-17746), and LNNano (Project TEM 24346).
© 2020 Elsevier Inc.
- AB toxin
- Poisonous egg defense