Pavel K. Kuzmichev

Personal information

2006 - 2015: MS and PhD student, junior research fellow in Laboratory of biomolecular NMR Spectroscopy  Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry RAS

2015: j.r.f., MIPT

Education

PeriodCountry, cityEducation institutionAdditional info
2009 Russia, Moscow Moscow Institute of Physics and Technology (State University) (MIPT), dep. molecular and biological physics PhD student
2003–2009 Russia, Moscow Moscow Institute of Physics and Technology (State University) (MIPT), dep. molecular and biological physics Ms in physical and chemical biology and biotechnology thesis: «Solubilization and refolding of G-coupled protein receptors»

Scientific interests

Bioorganic chemistry, molecular biology, biophysics, NMR, biological membranes, membrane proteins and membrane active peptides, G-coupled protein receptors, ion channels, nanotechnology.

Awards & honors

2008: Professor Yu.A. Ovchinnikov's scholarship

Selected publications

  1. Mineev K.S., Goncharuk S.A., Kuzmichev P.K., Vilar M., Arseniev A.S. (2015). NMR Dynamics of Transmembrane and Intracellular Domains of p75NTR in Lipid-Protein Nanodiscs. Biophys. J. 109 (4), 772–82 [+]

    P75NTR is a type I integral membrane protein that plays a key role in neurotrophin signaling. However, structural data for the receptor in various functional states are sparse and controversial. In this work, we studied the spatial structure and mobility of the transmembrane and intracellular parts of p75NTR, incorporated into lipid-protein nanodiscs of various sizes and compositions, by solution NMR spectroscopy. Our data reveal a high level of flexibility and disorder in the juxtamembrane chopper domain of p75NTR, which results in the motions of the receptor death domain being uncoupled from the motions of the transmembrane helix. Moreover, none of the intracellular domains of p75NTR demonstrated a propensity to interact with the membrane or to self-associate under the experimental conditions. The obtained data are discussed in the context of the receptor activation mechanism.

    ID:1312
  2. Gushchin I., Chervakov P., Kuzmichev P., Popov A.N., Round E., Borshchevskiy V., Ishchenko A., Petrovskaya L., Chupin V., Dolgikh D.A., Arseniev A.S., Kirpichnikov M., Gordeliy V. (2013). Structural insights into the proton pumping by unusual proteorhodopsin from nonmarine bacteria. Proc. Natl. Acad. Sci. U.S.A. 110 (31), 12631–6 [+]

    Light-driven proton pumps are present in many organisms. Here, we present a high-resolution structure of a proteorhodopsin from a permafrost bacterium, Exiguobacterium sibiricum rhodopsin (ESR). Contrary to the proton pumps of known structure, ESR possesses three unique features. First, ESR's proton donor is a lysine side chain that is situated very close to the bulk solvent. Second, the α-helical structure in the middle of the helix F is replaced by 3(10)- and π-helix-like elements that are stabilized by the Trp-154 and Asn-224 side chains. This feature is characteristic for the proteorhodopsin family of proteins. Third, the proton release region is connected to the bulk solvent by a chain of water molecules already in the ground state. Despite these peculiarities, the positions of water molecule and amino acid side chains in the immediate Schiff base vicinity are very well conserved. These features make ESR a very unusual proton pump. The presented structure sheds light on the large family of proteorhodopsins, for which structural information was not available previously.

    ID:1260
  3. Goncharuk S.A., Shulga A.A., Ermolyuk Y.S., Kuzmichev P.K., Sobol V.A., Bocharov E.V., Chupin V.V., Arseniev A.S., Kirpichnikov M.P. (2009). Bacterial synthesis, purification, and solubilization of membrane protein KCNE3, a regulator of voltage-gated potassium channels. Biochemistry Mosc. 74 (12), 1344–9 [+]

    An efficient method is described for production of membrane protein KCNE3 and its isotope labeled derivatives ((15)N-, (15)N-/13C-) in amounts sufficient for structural-functional investigations. The purified protein preparation within different detergent micelles was characterized using dynamic light scattering, CD spectroscopy, and NMR spectroscopy. It is shown that within DPC/LDAO micelles the protein is in monomeric form and acquires mainly alpha-helical conformation. The existence of cross-peaks for all glycines of the (15)N-HSQC NMR spectra as well as relatively small line widths (~20 Hz) confirm the high quality of the preparation and the possibility of obtaining structural-dynamic information on KCNE3 by high resolution heteronuclear NMR spectroscopy.

    ID:273