Laboratory of pharmacokinetics

NamePositionContacts
Dmitry Zinchenko, Ph.D.Head of lab.zdv@bibch.ru
Tatyana Muranova, Ph.D.s. r. f.muranova@bibch.ru
Alexey Danilkovich, Ph.D.r. f.danilkovich@bibch.ru+7(4967)73-08-93
Igor' Udobichenko, Ph.D.r. f.iudovichenko1@yandex.ru+7(926)5926321

Former members:

Valery Lipkin, corr. member of the RASheadvmlipkin@ibch.ru
Sergey Matveev, Ph.D.r. f.sergey.matveev@uky.edu

All publications (show selected)

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Dmitry Zinchenko

Caveolin-1 as a regulator of neuronal calcium sensor proteins in phototransduction system.

Caveolin-1 is the major regulatory protein of detergent resistant membranes (DRM), which associating with regulation of signaling activity in different organism systems. Rod cell membranes contain cholesterol-rich DRM, which accumulate visual cascade proteins. In this study, photoreceptor Ca2+-binding proteins recoverin, NCS1, GCAP1, and GCAP2, belonging to neuronal calcium sensor (NCS) family, were recognized as novel caveolin-1 interacting partners. We demonstrated that all studied proteins co-precipitate with -from rod outer segment membranes, and can directly interact with caveolin-1. Pull-down assay, surface plasmon resonance spectroscopy and isothermal titration calorimetry data indicate that there is interaction with full-length caveolin-1 N-terminal domain (1-101) and caveolin-1 scaffolding domain (81-101). Interestingly that this interaction occurs only in absence of calcium ions, what is supported by surface localization of caveolin-1 interaction site in Ca2+-free NCS proteins state. Caveolin-1 increase Ca2+-sensitivity of recoverin, and as a consequence, makes its inhibitory activity to rhodopsinkinase more pronounced, but not interfere with recoverin-­rhodopsinkinase interaction. Amount of free Ca2+, required for this process, consider caveolin­1 influence, become in good agreement with physiological conditions of photoreceptor cell. GCAP-2 is upregulated by caveolin-1 in Ca2+-free state, which increase guanylate cyclase activity. It seems that there is a common mechanism of interaction between caveolin-1 and NCS proteins in Ca2+-free state. For recoverin increasing of Ca2+-sensitivity is due to the stabilization of the open conformation of its second Ca2+-binding domain EF2. Obtained data suggest that at low calcium NCS proteins are compartmentalized in photoreceptor rafts via binding to caveolin-1, what enhances their activity or ensures their faster responses on Ca2+-signals.

Publications

  1. Vladimirov VI, Zernii EY, Baksheeva VE, Wimberg H, Kazakov AS, Tikhomirova NK, Nemashkalova EL, Mitkevich VA, Zamyatnin AA, Lipkin VM, Philippov PP, Permyakov SE, Senin II, Koch KW, Zinchenko DV (2018). Photoreceptor calcium sensor proteins in detergent-resistant membrane rafts are regulated via binding to caveolin-1. Cell Calcium 73, 55–69

Development of efficient solid-phase synthesis methods for the preparation of peptides that possess immunosuppresive activity

In collaboration with Laboratory of Biological Testing,  Group of Peptide Chemistry

The relative efficacies of several synthesis methods have been investigated aiming to prepare target peptides that are quite potent in a test for suppression of the experimental autoimmune encephalomyelitis. Toward this goal chemical yields of the target peptides as well as side product distributions in the samples of the peptides obtained using various methods have been evaluated and quantitatively characterized. A variation of Fmoc/tBu methodology have been found to be the most efficient providing targets peptides in the highest chemical yield since it allowed to use a broader range of  activated amino acid derivatives. Side products of the amino acid doubling were detected among the side products of chemical synthesis. Some of the amino acid doubling side products made the HPLC purification step to be complicated. Nevertheless the synthetic methods developed allowed the preparation of the target peptides in sufficient quantities.

Development of peptide drugs for the treatment of multiple sclerosis

In collaboration with Laboratory of hormonal regulation proteins,  Group of Peptide Chemistry,  Laboratory of Biological Testing

Multiple sclerosis is a chronic autoimmune disease with neurological pathology. The dominant role of immunological processes in the development of the disease dictates the need for medications that specifically minimize the activity of immune processes. The peptides A8AMS and mA8AMS, homologous to the fragment of the human IgG VH domain, have been shown to act in vitro and in vivo, effectively reducing the symptoms of experimental autoimmune encephalomyelitis, an animal model of multiple sclerosis. The results provide new opportunities for the development of peptide drugs for the treatment of multiple sclerosis.

Molecular mechanism of the non-opioid beta-endorphin action

In collaboration with Laboratory of hormonal regulation proteins,  Laboratory of toxicology in vitro

Using a synthetic peptide octarphin (TPLVTLFK), a selective agonist of non-opioid beta-endorphin receptor, data on the molecular mechanism of the non-opioid action of the hormone have been obtained. It was found that the non-opioid effect of beta- endorphin on the target cell is realized by the following way: increase in the inducible NO synthase expression → increase in NO production → increase in the activity of soluble guanylate cyclase → increase in the intracellular level of cGMP.