CC BY
7
Vestnik FEB RAS. 2018. № 6 Supplement
UDC 577.2 DOI: 10.25808/08697698.2018.202.6S.074
H.K. KIM, S.W. CHO, N.P. MISHCHENKO, E.A. VASILEVA, S.A. FEDOREYEV, V.A. STONIK, J.HAN
A novel atypical PKC-iota inhibitor, echinochrome A,
enhances cardiomyocyte differentiation from mouse embryonic stem cells
Key words: echinochrome A, cardiomyocyte differentiation, mitochondrial function, reactive oxygen species, stem cells, kinase activity
Echinochrome A (EchA) is a marine bioproduct extracted from sea urchins having antioxidant, antimicrobial, anti-inflammatory, and chelating effects which is the active component of the clinical drug histochrome. We investigated the potential use of Ech A for inducing cardiomyocyte differentiation from mouse embryonic stem cells (mESCs). We also assessed the effects of Ech A on mitochondrial mass, inner membrane potential (Aym), reactive oxygen species generation, and levels of Ca2+. To identify the direct target of Ech A, we performed in vitro kinase activity and surface plasmon resonance binding assays. Ech A dose-dependently enhanced cardiomyocyte differentiation with higher beating rates. Ech A (50 ^M) increased the mitochondrial mass and membrane potential but did not alter the mitochondrial superoxide and Ca2+ levels. The in vitro kinase activity of the atypical protein kinase C-iota (PKCi) was significantly decreased by 50 ^M of Ech A with an IC50 for PKCi activity of 107 ^M. Computational protein-ligand docking simulation results suggested the direct binding of Ech A to PKCi, and surface plasmon resonance confirmed the direct binding with a low KD of 6.3 nM. Therefore, Ech A is a potential drug for enhancing cardiomyocyte differentiation from mESCs through direct binding and inhibition of PKCi activity.
The work was supported by by grants from the Priority Research Centers Program, Basic Science Research Program, and International Research & Development Program through the National Research Foundation of Korea (NRF), funded by the Ministry of Education, Science, and Technology (2010-0020224,2015R1A2A1A13001900, 2015R1D1A1A01057937, and 2017K1A3A1A49070056). The study was carried out under support of the Ministry of Education and Science of the Russian Federation (RFMEFI61317X0076).
* KIM Hyoung Kyu - PhD, Associate Professor, CHO Sung Woo - MD, PhD, HAN Jin - MD, PhD Professor and Chair of Department of Physiology, Director of Cardiovascular and Metabolic Disease Center (National Research Laboratory for Mitochondrial Signaling, Department of Physiology, Department of Health Sciences and Technology, BK21 Plus Project Team, Cardiovascular and Metabolic Disease Center, Inje University College of Medicine, Busan, Korea); FEDOREEV Sergei Alexandrovich - DSc, Head of the Laboratory, MISHCHENKO Natalia Petrovna - PhD, Leading Researcher, VASILEVA Elena Andreevna - Junior Researcher, (G.B. Elyakov Pacific Institute of Bioorganic Chemistry, FEB RAS, Vladivostok, Russia). *E-mail: estrus74@gmail.com
REFERENCES:
1. Jeong S, Kim H, Song I-S, Lee S, Ko K, Rhee B, Kim N, Mishchenko N, Fedoryev S, Stonik V, Han J (2014) Echinochrome A Protects Mitochondrial Function in Cardiomyocytes against Cardiotoxic Drugs. Mar Drugs 12:29222936
2. Jeong S, Kim H, Song I-S, Noh S, Marquez J, Ko K, Rhee B, Kim N, Mishchenko N, Fedoreyev S, Stonik V, Han J (2014) Echinochrome A Increases Mitochondrial Mass and Function by Modulating Mitochondrial Biogenesis Regulatory Genes. Marine Drugs 12:4602-4615
3. Kim HK, Youm JB, Jeong SH, Lee SR, Song IS, Ko TH, Pronto JR, Ko KS, Rhee BD, Kim N, Nilius B, Mischchenko NP, Fedoreyev SA, Stonik VA, Han J (2015) Echinochrome A regulates phosphorylation of phospholamban Ser16 and Thr17 suppressing cardiac SERCA2A Ca(2)(+) reuptake. Pflugers Arch 467:2151-63.
