New Insectotoxin from Tibellus Oblongus Spider Venom Presents Novel Adaptation of ICK Fold
Yuliya Korolkova,
Ekaterina Maleeva,
Alexander Mikov,
Anna Lobas,
Elizaveta Solovyeva,
Mikhail Gorshkov,
Yaroslav Andreev,
Steve Peigneur,
Jan Tytgat,
Fedor Kornilov,
Vladislav Lushpa,
Konstantin Mineev,
Sergey Kozlov
Affiliations
Yuliya Korolkova
Department of Molecular Neurobiology, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry RAS, 16/10 Miklukho-Maklay Str., 117997 Moscow, Russia
Ekaterina Maleeva
Department of Molecular Neurobiology, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry RAS, 16/10 Miklukho-Maklay Str., 117997 Moscow, Russia
Alexander Mikov
Department of Molecular Neurobiology, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry RAS, 16/10 Miklukho-Maklay Str., 117997 Moscow, Russia
Anna Lobas
V.L. Talrose Institute for Energy Problems of Chemical Physics, N.N. Semenov Federal Research Center for Chemical Physics, Russian Academy of Sciences, 38 Leninsky Pr., Bld. 2, 119334 Moscow, Russia
Elizaveta Solovyeva
V.L. Talrose Institute for Energy Problems of Chemical Physics, N.N. Semenov Federal Research Center for Chemical Physics, Russian Academy of Sciences, 38 Leninsky Pr., Bld. 2, 119334 Moscow, Russia
Mikhail Gorshkov
V.L. Talrose Institute for Energy Problems of Chemical Physics, N.N. Semenov Federal Research Center for Chemical Physics, Russian Academy of Sciences, 38 Leninsky Pr., Bld. 2, 119334 Moscow, Russia
Yaroslav Andreev
Department of Molecular Neurobiology, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry RAS, 16/10 Miklukho-Maklay Str., 117997 Moscow, Russia
Steve Peigneur
Toxicology and Pharmacology, University of Leuven (KU Leuven), Campus Gasthuisberg, O&N2, Herestraat 49, P.O. Box 922, B-3000 Leuven, Belgium
Jan Tytgat
Toxicology and Pharmacology, University of Leuven (KU Leuven), Campus Gasthuisberg, O&N2, Herestraat 49, P.O. Box 922, B-3000 Leuven, Belgium
Fedor Kornilov
Department of Biological and Medicinal Physics, Moscow Institute of Physics and Technology (National Research University), 9 Institutsky Per., 141700 Dolgoprudny, Russia
Vladislav Lushpa
Department of Biological and Medicinal Physics, Moscow Institute of Physics and Technology (National Research University), 9 Institutsky Per., 141700 Dolgoprudny, Russia
Konstantin Mineev
Department of Biological and Medicinal Physics, Moscow Institute of Physics and Technology (National Research University), 9 Institutsky Per., 141700 Dolgoprudny, Russia
Sergey Kozlov
Department of Molecular Neurobiology, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry RAS, 16/10 Miklukho-Maklay Str., 117997 Moscow, Russia
The Tibellus oblongus spider is an active predator that does not spin webs and remains poorly investigated in terms of venom composition. Here, we present a new toxin, named Tbo-IT2, predicted by cDNA analysis of venom glands transcriptome. The presence of Tbo-IT2 in the venom was confirmed by proteomic analyses using the LC-MS and MS/MS techniques. The distinctive features of Tbo-IT2 are the low similarity of primary structure with known animal toxins and the unusual motif of 10 cysteine residues distribution. Recombinant Tbo-IT2 (rTbo-IT2), produced in E. coli using the thioredoxin fusion protein strategy, was structurally and functionally studied. rTbo-IT2 showed insecticidal activity on larvae of the housefly Musca domestica (LD100 200 μg/g) and no activity on the panel of expressed neuronal receptors and ion channels. The spatial structure of the peptide was determined in a water solution by NMR spectroscopy. The Tbo-IT2 structure is a new example of evolutionary adaptation of a well-known inhibitor cystine knot (ICK) fold to 5 disulfide bonds configuration, which determines additional conformational stability and gives opportunities for insectotoxicity and probably some other interesting features.
