{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:07:21Z","timestamp":1760144841026,"version":"build-2065373602"},"reference-count":52,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2024,5,27]],"date-time":"2024-05-27T00:00:00Z","timestamp":1716768000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100010269","name":"Wellcome Trust project","doi-asserted-by":"publisher","award":["GR074359","28\/B14085","BB\/D523078\/1"],"award-info":[{"award-number":["GR074359","28\/B14085","BB\/D523078\/1"]}],"id":[{"id":"10.13039\/100010269","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000268","name":"Biotechnology and Biological Science Research Council Core","doi-asserted-by":"publisher","award":["GR074359","28\/B14085","BB\/D523078\/1"],"award-info":[{"award-number":["GR074359","28\/B14085","BB\/D523078\/1"]}],"id":[{"id":"10.13039\/501100000268","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Medical Research Council (MRC) studentship","award":["GR074359","28\/B14085","BB\/D523078\/1"],"award-info":[{"award-number":["GR074359","28\/B14085","BB\/D523078\/1"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Toxins"],"abstract":"<jats:p>\u03b1-Latrotoxin (\u03b1-LTX) was found to form two-dimensional (2D) monolayer arrays in solution at relatively low concentrations (0.1 mg\/mL), with the toxin tetramer constituting a unit cell. The crystals were imaged using cryogenic electron microscopy (cryoEM), and image analysis yielded a ~12 \u00c5 projection map. At this resolution, no major conformational changes between the crystalline and solution states of \u03b1-LTX tetramers were observed. Electrophysiological studies showed that, under the conditions of crystallization, \u03b1-LTX simultaneously formed multiple channels in biological membranes that displayed coordinated gating. Two types of channels with conductance levels of 120 and 208 pS were identified. Furthermore, we observed two distinct tetramer conformations of tetramers both when observed as monodisperse single particles and within the 2D crystals, with pore diameters of 11 and 13.5 \u00c5, suggestive of a flickering pore in the middle of the tetramer, which may correspond to the two states of toxin channels with different conductance levels. We discuss the structural changes that occur in \u03b1-LTX tetramers in solution and propose a mechanism of \u03b1-LTX insertion into the membrane. The propensity of \u03b1-LTX tetramers to form 2D crystals may explain many features of \u03b1-LTX toxicology and suggest that other pore-forming toxins may also form arrays of channels to exert maximal toxic effect.<\/jats:p>","DOI":"10.3390\/toxins16060248","type":"journal-article","created":{"date-parts":[[2024,5,27]],"date-time":"2024-05-27T08:36:22Z","timestamp":1716798982000},"page":"248","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["\u03b1-Latrotoxin Tetramers Spontaneously Form Two-Dimensional Crystals in Solution and Coordinated Multi-Pore Assemblies in Biological Membranes"],"prefix":"10.3390","volume":"16","author":[{"given":"Alexis","family":"Rohou","sequence":"first","affiliation":[{"name":"Division of Cell and Molecular Biology, Imperial College London, Exhibition Road, London SW7 2AZ, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Edward P.","family":"Morris","sequence":"additional","affiliation":[{"name":"Division of Structural Biology, The Institute of Cancer Research (ICR), London SW7 3RP, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4497-3421","authenticated-orcid":false,"given":"Julia","family":"Makarova","sequence":"additional","affiliation":[{"name":"Faculty of Biology and Biotechnology, HSE University, 117997 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alexander G.","family":"Tonevitsky","sequence":"additional","affiliation":[{"name":"Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry RAS, 117997 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuri A.","family":"Ushkaryov","sequence":"additional","affiliation":[{"name":"Division of Cell and Molecular Biology, Imperial College London, Exhibition Road, London SW7 2AZ, UK"},{"name":"Medway School of Pharmacy, University of Kent, Chatham ME4 4TB, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,5,27]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1016\/0163-7258(89)90024-7","article-title":"\u03b1-Latrotoxin and Related Toxins","volume":"42","author":"Rosenthal","year":"1989","journal-title":"Pharmacol. 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