caa a22 4500 606234012 CHVBK 20210128101235.0 cr unu---uuuuu 210128e20150401xx s 000 0 eng 10.1007/s11010-014-2313-y doi (NATIONALLICENCE)springer-10.1007/s11010-014-2313-y Rutin stimulates sarcoplasmic reticulum Ca2+-ATPase activity (SERCA1) and protects SERCA1 from peroxynitrite mediated injury [Elektronische Daten] [Jana Viskupicova, Miriam Strosova, Petronela Zizkova, Magdalena Majekova, Lubica Horakova] In this study we analyzed the protective action of the flavonoid rutin on peroxynitrite (ONOO−) mediated impairment of sarcoplasmic reticulum Ca2+-ATPase (SERCA1 isoform), especially related to posttranslational and conformational changes. Rutin concentration dependently protected ONOO− induced SERCA1 activity decrease with effective concentration EC50 of 18±1.5µM. Upon treatment with ONOO−, this flavonoid also prevented SERCA1 from thiol group oxidation and significantly reduced tyrosine nitration and protein carbonyl formation. In the absence of ONOO−, rutin (250 and 350µM) stimulated SERCA1 activity at 2.1mM [ATP] and 10µM [Ca2+]free. According to changes in the kinetic parameters V max and K m with regard to [ATP], rutin (250µM) increased the rate of enzyme catalysis and decreased the affinity of SERCA1 to ATP. FITC fluorescence decreased in the presence of rutin (150 and 250µM), indicating conformational changes in the cytosolic ATP binding region of SERCA1. In silico study confirmed the binding of rutin in the cytosolic region of SERCA1, in the vicinity of the ATP binding site. Residue Glu183 localized within the conserved TGES loop was identified to play a key role in rutin-SERCA1 interaction (H-bond length of 1.7Å), elucidating the ability of rutin to affect the affinity of SERCA1 to ATP. The binding of rutin in the proximity of Lys515 is likely to cause a decrease in FITC fluorescence. Springer Science+Business Media New York, 2014 Rutin nationallicence SERCA1 nationallicence ONOO− nationallicence Structural and conformational changes nationallicence In silico study nationallicence Viskupicova Jana Department of Biochemical Pharmacology, Institute of Experimental Pharmacology and Toxicology, Slovak Academy of Sciences, 841 04, Bratislava, Slovakia aut Strosova Miriam Pharmacology, Pharmaceutical Sciences, University of Geneva, 1211, Geneva 4, Switzerland aut Zizkova Petronela Department of Biochemical Pharmacology, Institute of Experimental Pharmacology and Toxicology, Slovak Academy of Sciences, 841 04, Bratislava, Slovakia aut Majekova Magdalena Department of Biochemical Pharmacology, Institute of Experimental Pharmacology and Toxicology, Slovak Academy of Sciences, 841 04, Bratislava, Slovakia aut Horakova Lubica Department of Biochemical Pharmacology, Institute of Experimental Pharmacology and Toxicology, Slovak Academy of Sciences, 841 04, Bratislava, Slovakia aut Molecular and Cellular Biochemistry Springer US; http://www.springer-ny.com 402/1-2(2015-04-01), 51-62 0300-8177 402:1-2<51 2015 402 11010 https://doi.org/10.1007/s11010-014-2313-y text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s11010-014-2313-y text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Viskupicova Jana Department of Biochemical Pharmacology, Institute of Experimental Pharmacology and Toxicology, Slovak Academy of Sciences, 841 04, Bratislava, Slovakia aut NATIONALLICENCE 700 1- Strosova Miriam Pharmacology, Pharmaceutical Sciences, University of Geneva, 1211, Geneva 4, Switzerland aut NATIONALLICENCE 700 1- Zizkova Petronela Department of Biochemical Pharmacology, Institute of Experimental Pharmacology and Toxicology, Slovak Academy of Sciences, 841 04, Bratislava, Slovakia aut NATIONALLICENCE 700 1- Majekova Magdalena Department of Biochemical Pharmacology, Institute of Experimental Pharmacology and Toxicology, Slovak Academy of Sciences, 841 04, Bratislava, Slovakia aut NATIONALLICENCE 700 1- Horakova Lubica Department of Biochemical Pharmacology, Institute of Experimental Pharmacology and Toxicology, Slovak Academy of Sciences, 841 04, Bratislava, Slovakia aut NATIONALLICENCE 773 0- Molecular and Cellular Biochemistry Springer US; http://www.springer-ny.com 402/1-2(2015-04-01), 51-62 0300-8177 402:1-2<51 2015 402 11010