{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,29]],"date-time":"2026-03-29T14:45:21Z","timestamp":1774795521362,"version":"3.50.1"},"reference-count":67,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2023,6,3]],"date-time":"2023-06-03T00:00:00Z","timestamp":1685750400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100006769","name":"Russian Science Foundation","doi-asserted-by":"publisher","award":["22-14-00326"],"award-info":[{"award-number":["22-14-00326"]}],"id":[{"id":"10.13039\/501100006769","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Toxins"],"abstract":"<jats:p>Ph\u03b11\u03b2 (PnTx3\u20136) is a neurotoxin from the spider Phoneutria nigriventer venom, originally identified as an antagonist of two ion channels involved in nociception: N-type voltage-gated calcium channel (CaV2.2) and TRPA1. In animal models, Ph\u03b11\u03b2 administration reduces both acute and chronic pain. Here, we report the efficient bacterial expression system for the recombinant production of Ph\u03b11\u03b2 and its 15N-labeled analogue. Spatial structure and dynamics of Ph\u03b11\u03b2 were determined via NMR spectroscopy. The N-terminal domain (Ala1\u2013Ala40) contains the inhibitor cystine knot (ICK or knottin) motif, which is common to spider neurotoxins. The C-terminal \u03b1-helix (Asn41\u2013Cys52) stapled to ICK by two disulfides exhibits the \u00b5s\u2013ms time-scale fluctuations. The Ph\u03b11\u03b2 structure with the disulfide bond patterns Cys1\u20135, Cys2\u20137, Cys3\u201312, Cys4\u201310, Cys6\u201311, Cys8\u20139 is the first spider knottin with six disulfide bridges in one ICK domain, and is a good reference to other toxins from the ctenitoxin family. Ph\u03b11\u03b2 has a large hydrophobic region on its surface and demonstrates a moderate affinity for partially anionic lipid vesicles at low salt conditions. Surprisingly, 10 \u00b5M Ph\u03b11\u03b2 significantly increases the amplitude of diclofenac-evoked currents and does not affect the allyl isothiocyanate (AITC)-evoked currents through the rat TRPA1 channel expressed in Xenopus oocytes. Targeting several unrelated ion channels, membrane binding, and the modulation of TRPA1 channel activity allow for considering Ph\u03b11\u03b2 as a gating modifier toxin, probably interacting with S1\u2013S4 gating domains from a membrane-bound state.<\/jats:p>","DOI":"10.3390\/toxins15060378","type":"journal-article","created":{"date-parts":[[2023,6,5]],"date-time":"2023-06-05T02:18:29Z","timestamp":1685931509000},"page":"378","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Recombinant Production, NMR Solution Structure, and Membrane Interaction of the Ph\u03b11\u03b2 Toxin, a TRPA1 Modulator from the Brazilian Armed Spider Phoneutria nigriventer"],"prefix":"10.3390","volume":"15","author":[{"given":"Ekaterina N.","family":"Lyukmanova","sequence":"first","affiliation":[{"name":"Department of Biology, MSU-BIT Shenzhen University, Shenzhen 518172, China"},{"name":"Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 119997 Moscow, Russia"},{"name":"Phystech School of Biological and Medical Physics, Moscow Institute of Physics and Technology (State University), 141701 Dolgoprudny, Russia"},{"name":"Interdisciplinary Scientific and Educational School of Moscow University \u201cMolecular Technologies of the Living Systems and Synthetic Biology\u201d, Faculty of Biology, Lomonosov Moscow State University, 119234 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7884-3257","authenticated-orcid":false,"given":"Pavel A.","family":"Mironov","sequence":"additional","affiliation":[{"name":"Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 119997 Moscow, Russia"},{"name":"Interdisciplinary Scientific and Educational School of Moscow University \u201cMolecular Technologies of the Living Systems and Synthetic Biology\u201d, Faculty of Biology, Lomonosov Moscow State University, 119234 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dmitrii S.","family":"Kulbatskii","sequence":"additional","affiliation":[{"name":"Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 119997 Moscow, Russia"},{"name":"Phystech School of Biological and Medical Physics, Moscow Institute of Physics and Technology (State University), 141701 Dolgoprudny, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mikhail A.","family":"Shulepko","sequence":"additional","affiliation":[{"name":"Department of Biology, MSU-BIT Shenzhen University, Shenzhen 518172, China"},{"name":"Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 119997 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3614-560X","authenticated-orcid":false,"given":"Alexander S.","family":"Paramonov","sequence":"additional","affiliation":[{"name":"Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 119997 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Elizaveta M.","family":"Chernaya","sequence":"additional","affiliation":[{"name":"Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 119997 Moscow, Russia"},{"name":"National Research University Higher School of Economics, 101000 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yulia A.","family":"Logashina","sequence":"additional","affiliation":[{"name":"Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 119997 Moscow, Russia"},{"name":"Institute of Molecular Medicine, Sechenov First Moscow State Medical University, 119991 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7628-0980","authenticated-orcid":false,"given":"Yaroslav A.","family":"Andreev","sequence":"additional","affiliation":[{"name":"Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 119997 Moscow, Russia"},{"name":"Institute of Molecular Medicine, Sechenov First Moscow State Medical University, 119991 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mikhail P.","family":"Kirpichnikov","sequence":"additional","affiliation":[{"name":"Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 119997 Moscow, Russia"},{"name":"Interdisciplinary Scientific and Educational School of Moscow University \u201cMolecular Technologies of the Living Systems and Synthetic Biology\u201d, Faculty of Biology, Lomonosov Moscow State University, 119234 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1383-3522","authenticated-orcid":false,"given":"Zakhar O.","family":"Shenkarev","sequence":"additional","affiliation":[{"name":"Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 119997 Moscow, Russia"},{"name":"Phystech School of Biological and Medical Physics, Moscow Institute of Physics and Technology (State University), 141701 Dolgoprudny, Russia"},{"name":"International Tomography Center SB RAS, 630090 Novosibirsk, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2023,6,3]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"717","DOI":"10.1042\/bj20031860","article-title":"Diversity of Folds in Animal Toxins Acting on Ion Channels","volume":"378","author":"Mouhat","year":"2004","journal-title":"Biochem. 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