Цетлин Виктор Ионович

Премии и заслуги

  • Лауреат Государственной премии СССР
  • Лауреат премии Гумбольдта

Избранные публикации

  1. Kasheverov I.E., Chugunov A.O., Kudryavtsev D.S., Ivanov I.A., Zhmak M.N., Shelukhina I.V., Spirova E.N., Tabakmakher V.M., Zelepuga E.A., Efremov R.G., Tsetlin V.I. (2016). High-Affinity α-Conotoxin PnIA Analogs Designed on the Basis of the Protein Surface Topography Method. Sci Rep 6, 36848 [+]

    Despite some success for small molecules, elucidating structure-function relationships for biologically active peptides - the ligands for various targets in the organism - remains a great challenge and calls for the development of novel approaches. Some of us recently proposed the Protein Surface Topography (PST) approach, which benefits from a simplified representation of biomolecules' surface as projection maps, which enables the exposure of the structure-function dependencies. Here, we use PST to uncover the "activity pattern" in α-conotoxins - neuroactive peptides that effectively target nicotinic acetylcholine receptors (nAChRs). PST was applied in order to design several variants of the α-conotoxin PnIA, which were synthesized and thoroughly studied. Among the best was PnIA[R9, L10], which exhibits nanomolar affinity for the α7 nAChR, selectivity and a slow wash-out from this target. Importantly, these mutations could hardly be delineated by "standard" structure-based drug design. The proposed combination of PST with a set of experiments proved very efficient for the rational construction of new bioactive molecules.

    ID:1604
  2. Stepanov A., Belyy A., Kasheverov I., Rybinets A., Dronina M., Dyachenko I., Murashev A., Knorre V., Sakharov D., Ponomarenko N., Tsetlin V., Tonevitsky A., Deyev S., Belogurov A., Gabibov A. (2016). Development of a recombinant immunotoxin for the immunotherapy of autoreactive lymphocytes expressing MOG-specific BCRs. Biotechnol. Lett. , [+]

    Myelin oligodendrocyte glycoprotein (MOG) is one of the major autoantigens in multiple sclerosis (MS), therefore selective depletion of autoreactive lymphocytes exposing MOG-specific B cell receptors (BCRs) would be beneficial in terms of MS treatment.

    ID:1524
  3. Lyukmanova E.N., Shulepko M.A., Kudryavtsev D., Bychkov M.L., Kulbatskii D.S., Kasheverov I.E., Astapova M.V., Feofanov A.V., Thomsen M.S., Mikkelsen J.D., Shenkarev Z.O., Tsetlin V.I., Dolgikh D.A., Kirpichnikov M.P. (2016). Human Secreted Ly-6/uPAR Related Protein-1 (SLURP-1) Is a Selective Allosteric Antagonist of α7 Nicotinic Acetylcholine Receptor. PLoS ONE 11 (2), e0149733 [+]

    SLURP-1 is a secreted toxin-like Ly-6/uPAR protein found in epithelium, sensory neurons and immune cells. Point mutations in the slurp-1 gene cause the autosomal inflammation skin disease Mal de Meleda. SLURP-1 is considered an autocrine/paracrine hormone that regulates growth and differentiation of keratinocytes and controls inflammation and malignant cell transformation. The majority of previous studies of SLURP-1 have been made using fusion constructs containing, in addition to the native protein, extra polypeptide sequences. Here we describe the activity and pharmacological profile of a recombinant analogue of human SLURP-1 (rSLURP-1) differing from the native protein only by one additional N-terminal Met residue. rSLURP-1 significantly inhibited proliferation (up to ~ 40%, EC50 ~ 4 nM) of human oral keratinocytes (Het-1A cells). Application of mecamylamine and atropine,-non-selective inhibitors of nicotinic acetylcholine receptors (nAChRs) and muscarinic acetylcholine receptors, respectively, and anti-α7-nAChRs antibodies revealed α7 type nAChRs as an rSLURP-1 target in keratinocytes. Using affinity purification from human cortical extracts, we confirmed that rSLURP-1 binds selectively to the α7-nAChRs. Exposure of Xenopus oocytes expressing α7-nAChRs to rSLURP-1 caused a significant non-competitive inhibition of the response to acetylcholine (up to ~ 70%, IC50 ~ 1 μM). It was shown that rSLURP-1 binds to α7-nAChRs overexpressed in GH4Cl cells, but does not compete with 125I-α-bungarotoxin for binding to the receptor. These findings imply an allosteric antagonist-like mode of SLURP-1 interaction with α7-nAChRs outside the classical ligand-binding site. Contrary to rSLURP-1, other inhibitors of α7-nAChRs (mecamylamine, α-bungarotoxin and Lynx1) did not suppress the proliferation of keratinocytes. Moreover, the co-application of α-bungarotoxin with rSLURP-1 did not influence antiproliferative activity of the latter. This supports the hypothesis that the antiproliferative activity of SLURP-1 is related to 'metabotropic' signaling pathway through α7-nAChR, that activates intracellular signaling cascades without opening the receptor channel.

    ID:1420
  4. Lyukmanova E.N., Shenkarev Z.O., Shulepko M.A., Paramonov A.S., Chugunov A.O., Janickova H., Dolejsi E., Dolezal V., Utkin Y.N., Tsetlin V.I., Arseniev A.S., Efremov R.G., Dolgikh D.A., Kirpichnikov M.P. (2015). Structural Insight into Specificity of Interactions between Nonconventional Three-finger Weak Toxin from Naja kaouthia (WTX) and Muscarinic Acetylcholine Receptors. J. Biol. Chem. 290 (39), 23616–30 [+]

    Weak toxin from Naja kaouthia (WTX) belongs to the group of nonconventional "three-finger" snake neurotoxins. It irreversibly inhibits nicotinic acetylcholine receptors and allosterically interacts with muscarinic acetylcholine receptors (mAChRs). Using site-directed mutagenesis, NMR spectroscopy, and computer modeling, we investigated the recombinant mutant WTX analogue (rWTX) which, compared with the native toxin, has an additional N-terminal methionine residue. In comparison with the wild-type toxin, rWTX demonstrated an altered pharmacological profile, decreased binding of orthosteric antagonist N-methylscopolamine to human M1- and M2-mAChRs, and increased antagonist binding to M3-mAChR. Positively charged arginine residues located in the flexible loop II were found to be crucial for rWTX interactions with all types of mAChR. Computer modeling suggested that the rWTX loop II protrudes to the M1-mAChR allosteric ligand-binding site blocking the entrance to the orthosteric site. In contrast, toxin interacts with M3-mAChR by loop II without penetration into the allosteric site. Data obtained provide new structural insight into the target-specific allosteric regulation of mAChRs by "three-finger" snake neurotoxins.

    ID:1394
  5. Kasheverov I.E., Kudryavtsev D.S., Ivanov I.A., Zhmak M.N., Chugunov A.O., Tabakmakher V.M., Zelepuga E.A., Efremov R.G., Tsetlin V.I. (2015). Rational design of new ligands for nicotinic receptors on the basis of α-conotoxin PnIA. Dokl. Biochem. Biophys. 461, 106–9 [+]

    A variety of different subtypes of nicotinic acetylcholine receptors (nAChRs) and their involvement in a number of diseases and pathologies (Parkinson’s and Alzheimer’s diseases, schizophrenia, myasthenia, nicotine addiction) dictates the needs in potent and selective ligands for each subtype. These ligands can be used as a tool for detection and characterization of the distinct nAChR subtypes, as well as be the basis for drug design. Novel cholinergic ligands can emerge in the result of search among natural sources or design (with the use of modern computer modeling) on the basis of known molecules. The significance of the first way was confirmed in our hands by the detection of affinities of a set of marine alkaloids from sponges and ascidians towards some receptor subtypes. The most active of them — makaluvamines — showed micromolar affinity for muscle and neuronal α7 nAChRs. Application of the recently presented Protein Surface Topography method to known natural antagonist of some neuronal nAChRs — α-conotoxin PnIA — resulted in design of new potent analogs with nanomolar affinities for α7 nAChR. Radioactive derivatives of these analogs were successfully applied in radioligand tests for characterization of novel compounds and could be perspective as well for detection of α7 nAChR in the various preparations. Combining these two ways (search and design) was demonstrated in the synthesis of small peptide compounds on the basis of discovered by us in venom of Burmese Viper linear peptide azemiopsin — powerful blocker of muscle-type nAChRs. Some designed peptides have retained a certain affinity to receptor and showed high practical potential: in the absence of toxicity they contributed to the reduction of facial wrinkles (patent application for cosmetic use RU2013102410; PCT/RU2014/000032).

    ID:1393
  6. Hoang A.N., Vo H.D., Vo N.P., Kudryashova K.S., Nekrasova O.V., Feofanov A.V., Kirpichnikov M.P., Andreeva T.V., Serebryakova M.V., Tsetlin V.I., Utkin Y.N. (2014). Vietnamese Heterometrus laoticus scorpion venom: evidence for analgesic and anti-inflammatory activity and isolation of new polypeptide toxin acting on Kv1.3 potassium channel. Toxicon 77, 40–8 [+]

    The scorpion Heterometrus laoticus (Scorpionidae) inhabits Indochinese peninsula and is widely distributed in South-West Vietnam. Since no human fatalities caused by H. laoticus stings were reported, no systematic characterization of the venom was earlier done. In this study we report on biological activity of the venom from H. laoticus caught in Vietnamese province An Giang. The venom manifested a very low acute toxicity with LD50 of about 190 mg/kg body weight in mice at subcutaneous (s.c.) injection and 12 mg/kg at intravenous injection. The venom analgesic effects using tail immersion and writhing tests as well as anti-inflammatory effect using carrageenan test were analyzed at doses of 9.5 and 19 mg/kg at s.c. injections. It was found that at two doses tested H. laoticus venom showed both anti-nociceptive and anti-inflammatory activity. The venom was fractionated by means of gel-filtration and reversed-phase HPLC. As a result several polypeptide toxins were isolated and new toxin hetlaxin was identified. Its amino acid sequence was determined and binding to the extracellular vestibule of the K⁺-conducting pore of Kv1.1 and Kv1.3 potassium channels was studied. Hetlaxin belongs to the scorpion alpha-toxin family and is the first toxin isolated from H. laoticus venom which possesses high affinity (K(i) 59 nM) to Kv1.3 potassium channel.

    ID:1082
  7. Kasheverov I.E., Zhmak M.N., Fish A., Rucktooa P., Khruschov A.Y., Osipov A.V., Ziganshin R.H., D'hoedt D., Bertrand D., Sixma T.K., Smit A.B., Tsetlin V.I. (2009). Interaction of alpha-conotoxin ImII and its analogs with nicotinic receptors and acetylcholine-binding proteins: additional binding sites on Torpedo receptor. J. Neurochem. 111 (4), 934–44 [+]

    Изучение необычного по своим свойствам a-конотоксина ImII и его новых аналогов различными методами показало существование на природном холинорецепторе дополнительного участка связывания для этих пептидов отличного от сайта связывания «классических» агонистов и конкурентных антагонистов.

    ID:191
  8. Mordvintsev D.Y., Polyak Y.L., Rodionov D.I., Jakubik J., Dolezal V., Karlsson E., Tsetlin V.I., Utkin Y.N. (2009). Weak toxin WTX from Naja kaouthia cobra venom interacts with both nicotinic and muscarinic acetylcholine receptors. FEBS J. 276 (18), 5065–75 [+]

    В этой работе показано, что слабый нейротоксин WTX может взаимодействовать как с никотиновыми, так и с мускариновыми холинорецепторами. При этом в никотиновых холинорецепторах взаимодействует с ортостерическим участком связывания, а в мускариновых холинорецепторам – с аллостерическим участком.

    ID:193
  9. Osipov A.V., Kasheverov I.E., Makarova Y.V., Starkov V.G., Vorontsova O.V., Ziganshin R.K., Andreeva T.V., Serebryakova M.V., Benoit A., Hogg R.C., Bertrand D., Tsetlin V.I., Utkin Y.N. (2008). Naturally occurring disulfide-bound dimers of three-fingered toxins: a paradigm for biological activity diversification. J. Biol. Chem. 283 (21), 14571–80 [+]

    Впервые обнаружены трех-петельные токсины нового структурного типа: соединенные дисульфидами димеры кобратоксина с цитотоксинами, а также гомодимер кобратоксина. Показано, что в результате димеризации кобратоксин приобретает способность взаимодействовать с еще одним типом никотинового холинорецептора. Такая пост-трансляционная модификация может рассматриваться как еще один путь диверсификации биологической активности трех-петельных токсинов.

    ID:105
  10. Kasheverov I.E., Zhmak M.N., Vulfius C.A., Gorbacheva E.V., Mordvintsev D.Y., Utkin Y.N., van Elk R., Smit A.B., Tsetlin V.I. (2006). Alpha-conotoxin analogs with additional positive charge show increased selectivity towards Torpedo californica and some neuronal subtypes of nicotinic acetylcholine receptors. FEBS J. 273 (19), 4470–81 [+]

    В данной работе описан синтез и характеристика большой серии новых аналогов нескольких a-конотоксинов, содержащих замены ряда остатков на заряженные или противоположно заряженные. Некоторые из этих аналогов оказались значительно более эффективными лигандами соответствующих холинорецепторов, чем природные пептиды. Предложены компьютерные модели комплексов этих аналогов с рецепторами, объясняющие подобный эффект.

    ID:190
  11. Kasheverov I.E., Chiara D.C., Zhmak M.N., Maslennikov I.V., Pashkov V.S., Arseniev A.S., Utkin Y.N., Cohen J.B., Tsetlin V.I. (2006). alpha-Conotoxin GI benzoylphenylalanine derivatives. (1)H-NMR structures and photoaffinity labeling of the Torpedo californica nicotinic acetylcholine receptor. FEBS J. 273 (7), 1373–88 [+]

     

    С использованием фотоактивируемого производного одного из конотоксинов методом фотоаффинной модификации проведено частичное картирование лиганд-связывающего участка природного холинорецептора. Исходя из полученных данных, компьютерным моделированием предположена возможность двух ориентаций молекулы пептида в этом участке.

     

    ID:189
  12. Celie P.H., Kasheverov I.E., Mordvintsev D.Y., Hogg R.C., van Nierop P., van Elk R., van Rossum-Fikkert S.E., Zhmak M.N., Bertrand D., Tsetlin V.I., Sixma T.K., Smit A.B. (2005). Crystal structure of nicotinic acetylcholine receptor homolog AChBP in complex with an alpha-conotoxin PnIA variant. Nat. Struct. Mol. Biol. 12 (7), 582–8 [+]

    Впервые установлена кристаллическая структура ацетилхолин-связывающего белка в комплексе с контоксином. Полученные данные позволили охарактеризовать основные структурные особенности взаимодействия конотоксинов с холинорецепторами.

    ID:104
  13. Osipov A.V., Astapova M.V., Tsetlin V.I., Utkin Y.N. (2004). The first representative of glycosylated three-fingered toxins. Cytotoxin from the Naja kaouthia cobra venom. Eur. J. Biochem. 271 (10), 2018–27 [+]

    В этой работе впервые был описан гликозилированный цитотоксин, выделенный из яда кобры Naja kaouthia. Это первый представитель обширного семейства трех-петельных токсинов, для которого обнаружена пост-трансляционная модификация

    ID:192
  14. Utkin Y.N., Kukhtina V.V., Kryukova E.V., Chiodini F., Bertrand D., Methfessel C., Tsetlin V.I. (2001). "Weak toxin" from Naja kaouthia is a nontoxic antagonist of alpha 7 and muscle-type nicotinic acetylcholine receptors. J. Biol. Chem. 276 (19), 15810–5 [+]

    Эта работа показала, что давно известные «слабые» токсины способны взаимодействовать с никотиновыми холинорецепторами. При этом, обладая слабым сродством к рецептору, такие токсины связываются практически необратимо.

    ID:103
  15. Machold J., Utkin Y., Kirsch D., Kaufmann R., Tsetlin V., Hucho F. (1995). Photolabeling reveals the proximity of the alpha-neurotoxin binding site to the M2 helix of the ion channel in the nicotinic acetylcholine receptor. Proc. Natl. Acad. Sci. U.S.A. 92 (16), 7282–6 [+]

    Впервые локализован аминокислотный остаток холинорецептора, меченный фотоактивированным производным нейротоксина. Это позволило установить расположение молекулы токсина на рецепторе. Согласно полученным данным молекула токсина находится гораздо ближе к поверхности мембраны, чем считалось ранее.

    ID:102
  16. Kreienkamp H.J., Utkin Y.N., Weise C., Machold J., Tsetlin V.I., Hucho F. (1992). Investigation of ligand-binding sites of the acetylcholine receptor using photoactivatable derivatives of neurotoxin II from Naja naja oxiana. Biochemistry 31 (35), 8239–44 [+]

    В данной работе впервые с использованием фотоактивируемых производных нейротоксина установлено, какие субъединицы холинорецептора мышечного типа участвуют в формировании участка связывания агонистов/конкурентных антагонистов.

    ID:101