Structural basis of molecular mechanisms of signal transduction by the type I integral membrane proteins

Establishment of the relationship between structural organization of proteins and peculiarities of their functioning is one of the major problems of the contemporary science. At this, the special interest of investigators is attracted by the group of cell receptors, belonging to the type I integral membrane proteins. This group includes growth factor receptors, cytokine receptors, toll-like receptors and many others. Dysfunction of signaling systems of the specified proteins can result in the development of cancer, osteomyopathy, diabetes, autoimmune and neurodegenerative diseases. These proteins are interesting for both scientific laboratories and pharmaceutical production companies as specific targets for therapeutic agents. Although a significant portion of data can be found in the literature regarding the structural organization of extracellular and intracellular domains of type I membrane proteins in both active and inactive states, for the vast majority of such proteins there is no data available about the structure of their membrane domains and about their role in the process of transmembrane signal transduction. Another question that is still open is the way, how the interconnection between the structural states of different domains of type I membrane proteins is accomplished. To achieve the deeper understanding of the structural basis underlying the molecular mechanisms of signal transduction by type I membrane proteins one needs to first conduct the complex structural and biological study of their membrane domains, and, second, come from the structural investigations of isolated domains to the junctions of transmembrane and intracellular or transmembrane and extracellular domains.

In the frame of the present Project it is suggested to investigate the structural organization of dimeric transmembrane domains of the number of type I integral membrane proteins (among others from the TLR and Trk family), to study the effects of biologically relevant mutations on the structural and thermodynamic parameters of the dimerization of transmembrane domains of some receptors, to obtain the information about the influence of cytoplasmic juxtamembrane regions on the spatial structure of the transmembrane domains of the HER receptors. In addition, the influence of phosphorylation on the dynamic parameters of intrinsically disordered cytoplasmic domains of two-component signaling subunit CD79ab of B-cell receptor will be investigated. Moreover, by means of solution NMR spectroscopy we intend to study the spatial structure and dynamic parameters of the full-size type I membrane protein, NRADD, and of the construct containing transmembrane and intracellular domains of the neurotrophine receptor trkA. Results that are planned to be obtained in the course of the Project will allow to answer several questions, concerning the activation processes of the type I integral membrane proteins and to understand the principles of functioning of many signaling systems.

May 1, 2014 — December 31, 2018

Mineev K.S. (PI), Bocharova O.V. (Co-PI), Goncharuk S.A. (Co-PI), Nadezhdin K.D. (Co-PI), Artem'eva L.E., Bocharov E.V., Bragin P.E., Goncharuk M.V., Lesovoy D.M., Nol'de S.B.

Laboratory of biomolecular NMR-spectroscopy

Grant, RSF

List of publications

  1. Bragin PE, Mineev KS, Bocharova OV, Volynsky PE, Bocharov EV, Arseniev AS (2016). HER2 Transmembrane Domain Dimerization Coupled with Self-Association of Membrane-Embedded Cytoplasmic Juxtamembrane Regions. J Mol Biol 428 (1), 52–61
  2. Bocharov EV, Lesovoy DM, Pavlov KV, Pustovalova YE, Bocharova OV, Arseniev AS (2016). Alternative packing of EGFR transmembrane domain suggests that protein-lipid interactions underlie signal conduction across membrane. BIOCHIM BIOPHYS ACTA 1858 (6), 1254–1261
  3. Bocharov EV, Mineev KS, Pavlov KV, Akimov SA, Kuznetsov AS, Efremov RG, Arseniev AS (2017). Helix-helix interactions in membrane domains of bitopic proteins: Specificity and role of lipid environment. BIOCHIM BIOPHYS ACTA 1859 (4), 561–576
  4. Mineev KS, Nadezhdin KD (2017). Membrane mimetics for solution NMR studies of membrane proteins. Nanotechnol Rev 6 (1), 15–32
  5. Mineev KS, Nadezhdin KD, Goncharuk SA, Arseniev AS (2017). Façade detergents as bicelle rim-forming agents for solution NMR spectroscopy. Nanotechnol Rev 6 (1), 93–103
  6. Mineev KS, Nadezhdin KD, Goncharuk SA, Arseniev AS (2016). Characterization of Small Isotropic Bicelles with Various Compositions. Langmuir 32 (26), 6624–6637
  7. Mineev KS, Goncharuk SA, Goncharuk MV, Volynsky PE, Novikova EV, Aresinev AS (2017). Spatial structure of TLR4 transmembrane domain in bicelles provides the insight into the receptor activation mechanism. Sci Rep 7 (1), 6864
  8. Lesovoy DM, Mineev KS, Bragin PE, Bocharova OV, Bocharov EV, Arseniev AS (2017). NMR relaxation parameters of methyl groups as a tool to map the interfaces of helix–helix interactions in membrane proteins. J Biomol NMR 69 (3), 165–179
  9. Nadezhdin KD, Goncharuk SA, Arseniev AS, Mineev KS (2019). NMR structure of a full-length single-pass membrane protein NRADD. Proteins 87 (9), 786–790
  10. Goncharuk SA, Artemieva LE, Tabakmakher VM, Arseniev AS, Mineev KS (2018). CARD domain of rat RIP2 kinase: Refolding, solution structure, pH-dependent behavior and protein-protein interactions. PLoS One 13 (10), e0206244
  11. Kot EF, Goncharuk SA, Arseniev AS, Mineev KS (2018). Phase Transitions in Small Isotropic Bicelles. Langmuir 34 (11), 3426–3437
  12. Kot EF, Arseniev AS, Mineev KS (2018). Behavior of Most Widely Spread Lipids in Isotropic Bicelles. Langmuir 34 (28), 8302–8313
  13. Bragin PE, Kuznetsov AS, Bocharova OV, Volynsky PE, Arseniev AS, Efremov RG, Mineev KS (2018). Probing the effect of membrane contents on transmembrane protein-protein interaction using solution NMR and computer simulations. BIOCHIM BIOPHYS ACTA 1860 (12), 2486–2498
  14. Mineev KS, Goncharuk SA, Arseniev AS (2014). Toll-like receptor 3 transmembrane domain is able to perform various homotypic interactions: An NMR structural study. FEBS Lett 588 (21), 3802–3807
  15. Mineev KS, Panova SV, Bocharova OV, Bocharov EV, Arseniev AS (2015). The Membrane Mimetic Affects the Spatial Structure and Mobility of EGFR Transmembrane and Juxtamembrane Domains. Biochemistry 54 (41), 6295–6298