Жмак Максим Нургаянович

Кандидат химических наук

Научный сотрудник (Лаборатория лиганд-рецепторных взаимодействий)

Тел.: +7 (495) 330-73-74

Эл. почта: mzhmak@gmail.com

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.

  2. Kudryavtsev D.S., Shelukhina I.V., Son L.V., Ojomoko L.O., Kryukova E.V., Lyukmanova E.N., Zhmak M.N., Dolgikh D.A., Ivanov I.A., Kasheverov I.E., Starkov V.G., Ramerstorfer J., Sieghart W., Tsetlin V.I., Utkin Y.N. (2015). Neurotoxins from Snake Venoms and α-Conotoxin ImI Inhibit Functionally Active Ionotropic GABA Receptors. J. Biol. Chem. , [+]

    Ionotropic receptors of γ-aminobutyric acid (GABAAR) regulate neuronal inhibition and are targeted by benzodiazepines and general anesthetics. We show that a fluorescent derivative of α-cobratoxin (α-Ctx), belonging to the family of three-finger toxins (TFTs) from snake venoms, specifically stained the α1β3γ2 receptor; at 10 μM α-Ctx completely blocked GABA-induced currents in this receptor expressed in Xenopus oocytes (IC50 = 236 nM) and less potently inhibited α1β2γ2 ≈ α2β2γ2 > α5β2γ2 > α2β3γ2 and α1β3δ GABAARs. The α1β3γ2 receptor was also inhibited by some other TFTs: long α-neurotoxin Ls III and non-conventional toxin WTX. α-Conotoxin ImI displayed inhibitory activity as well. Electrophysiology experiments showed mixed competitive and non-competitive α-Ctx action. Fluorescent α-Ctx, however, could be displaced by muscimol indicating that most of the α-Ctx binding sites overlap with the orthosteric sites at the β/α subunit interface. Modeling and molecular dynamic studies indicated that α-Ctx or α-bungarotoxin seem to interact with GABAAR in a way similar to their interaction with the acetylcholine-binding protein or the ligand-binding domain of nicotinic receptors. This was supported by mutagenesis studies and experiments with α-conotoxin ImI and a chimeric Naja oxiana α-neurotoxin indicating that the major role in α-Ctx binding to GABAAR is played by the tip of its central loop II accomodating under loop C of the receptors.

  3. 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).

  4. Salnikov E.S., Aisenbrey C., Balandin S.V., Zhmak M.N., Ovchinnikova T.V., Bechinger B. (2011). Structure and alignment of the membrane-associated antimicrobial peptide arenicin by oriented solid-state NMR spectroscopy. Biochemistry 50 (18), 3784–95 [+]

    The antimicrobial arenicin peptides are cationic amphipathic sequences that strongly interact with membranes. Through a cystine ring closure a cyclic β-sheet structure is formed in aqueous solution, which persists when interacting with model membranes. In order to investigate the conformation, interactions, dynamics, and topology of their bilayer-associated states, arenicin 1 and 2 were prepared by chemical solid-phase peptide synthesis or by bacterial overexpression, labeled selectively or uniformly with (15)N, reconstituted into oriented membranes, and investigated by proton-decoupled (31)P and (15)N solid-state NMR spectroscopy. Whereas the (31)P NMR spectra indicate that the peptide induces orientational disorder at the level of the phospholipid head groups, the (15)N chemical shift spectra agree well with a regular β-sheet conformation such as the one observed in micellar environments. In contrast, the data do not fit the twisted β-sheet structure found in aqueous buffer. Furthermore, the chemical shift distribution is indicative of considerable conformational and/or topological heterogeneity when at the same time the (15)N NMR spectra exclude alignments of the peptide where the β-sheet lies side ways on the membrane surface. The ensemble of experimental constraints, the amphipathic character of the peptide, and in particular the distribution of the six arginine residues are in agreement with a boatlike dimer structure, similar or related to the one observed in micellar solution, that floats on the membrane surface with the possibility to oligomerize into higher order structures and/or to insert in a transmembrane fashion.

  5. Macháň R., Hof M., Chernovets T., Zhmak M.N., Ovchinnikova T.V., Sýkora J. (2011). Formation of arenicin-1 microdomains in bilayers and their specific lipid interaction revealed by Z-scan FCS. Analytical and bioanalytical chemistry , [+]

    Z-scan fluorescence correlation spectroscopy (FCS) is employed to characterize the interaction between arenicin-1 and supported lipid bilayers (SLBs) of different compositions. Lipid analogue C8-BODIPY 500/510C5-HPC and ATTO 465 labelled arenicin-1 are used to detect changes in lipid and peptide diffusion upon addition of unlabelled arenicin-1 to SLBs. Arenicin-1 decreases lipid mobility in negatively charged SLBs. According to diffusion law analysis, microdomains of significantly lower lipid mobility are formed. The analysis of peptide FCS data confirms the presence of microdomains for anionic SLBs. No indications of microdomain formation are detected in SLBs composed purely of zwitterionic lipids. Additionally, our FCS results imply that arenicin-1 exists in the form of oligomers and/or aggregates when interacting with membranes of both compositions.

  6. 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.

  7. Ovchinnikova T.V., Shenkarev Z.O., Nadezhdin K.D., Balandin S.V., Zhmak M.N., Kudelina I.A., Finkina E.I., Kokryakov V.N., Arseniev A.S. (2007). Recombinant expression, synthesis, purification, and solution structure of arenicin. Biochem. Biophys. Res. Commun. 360 (1), 156–62 [+]

    Arenicins are 21-residue cationic antimicrobial peptides, isolated from marine polychaeta Arenicola marina. In order to determine a high-resolution three-dimensional structure of arenicin-2, the recombinant peptide was overexpressed as a fused form in Escherichia coli. Both arenicin isoforms were synthesized using the Fmoc-based solid-phase strategy. Recombinant and synthetic arenicins were purified, and their antimicrobial and spectroscopic properties were analyzed. NMR investigation shows that in water solution arenicin-2 displays a prolonged beta-hairpin, formed by two antiparallel beta-strands and stabilized by one disulfide and nine hydrogen bonds. A significant right-handed twist in the beta-sheet is deprived the peptide surface of amphipathicity. CD spectroscopic analysis indicates that arenicin-2 binds to the SDS and DPC micelles, and conformation of the peptide is significantly changed upon binding. Arenicin strongly binds to anionic lipid (POPE/POPG) vesicles in contrast with zwitterionic (POPC) ones. These results suggest that arenicins are membrane active peptides and point to possible mechanism of their selectivity toward bacterial cells.

  8. 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.

  9. 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.


  10. 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.

  11. Volpina O.M., Surovoy A.Y., Zhmak M.N., Kuprianova M.A., Koroev D.O., Chepurkin A.V., Toloknov A.S., Ivanov V.T. (1999). A peptide construct containing B-cell and T-cell epitopes from the foot-and-mouth disease viral VP1 protein induces efficient antiviral protection. Vaccine 17 (6), 577–84 [+]

    Новая пептидная конструкция Palm135-158-GGA-170-188(Acm) содержит специфический T-хелперный эпитоп последовательносчти 170—188 белка VP1и основной антигенный район 135—158 белка VP1 вируса ящура. Конструкция проявляет более высокую протективную, антигенную, иммуногенную активность, а также способность к Т-клеточной пролиферации, чем ранее описанный пептид Palm(2)135—159.

  12. Volpina O.M., Yarov A.V., Zhmak M.N., Kuprianova M.A., Chepurkin A.V., Toloknov A.S., Ivanov V.T. (1996). Synthetic vaccine against foot-and-mouth disease based on a palmitoyl derivative of the VP1 protein 135-159 fragment of the A22 virus strain. Vaccine 14 (14), 1375–80 [+]

    Синетезирован пептид Palm2 135—159, представляющий собой дипальмитоильное производное фрагмента 135—159 белка VP1 вируса ящура штамма A22. Показано, что пептид Palm2 135—159 проявляет противовирусную активность на мышах, морских свинках и овцах. Однократное введение синтетической пептидной вакцины обеспечивает защиту овец от заболевания ящуром в течение 1 год. Пептидная вакцина разрешена к применению на территории России.