Rutin stimulates sarcoplasmic reticulum Ca2+-ATPase activity (SERCA1) and protects SERCA1 from peroxynitrite mediated injury

Abstract

In this study we analyzed the protective action of the flavonoid rutin on peroxynitrite (ONOO(-)) mediated impairment of sarcoplasmic reticulum Ca(2+)-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 ± 1.5 µM. 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 350 µM) stimulated SERCA1 activity at 2.1 mM [ATP] and 10 µM [Ca(2+)]free. According to changes in the kinetic parameters V max and K m with regard to [ATP], rutin (250 µM) increased the rate of enzyme catalysis and decreased the affinity of SERCA1 to ATP. FITC fluorescence decreased in the presence of rutin (150 and 250 µM), 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 1.7 Å), 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.

Rutin Modulates MAPK Pathway Differently from Quercetin in Angiotensin II-Induced H9c2 Cardiomyocyte Hypertrophy

“…Flavonoids have been reported to increase SOD activity by binding their hydroxyl group to the SOD molecules, thereby stabilizing them [ 26 , 27 ]. Both compounds are able to scavenge superoxide and peroxynitrite radicals [ 12 , 28 , 29 ]. However, rutin had a lower ability than quercetin in augmenting SOD activity, which could be due to the difference in their structures, which warrants further investigation.…”

Caloric restriction induces energy-sparing alterations in skeletal muscle contraction, fiber composition and local thyroid hormone metabolism that persist during catch-up fat upon refeeding

“…The enzymatic activity of the SERCA pumps was measured by an NADH-coupled assay (Warren et al, 1974 ; Strosova et al, 2011 ; Viskupicova et al, 2015 ). Muscle microsomes were prepared from the gastrocnemius muscle of semistarved, refed and controls rats according to Warren et al ( 1974 ), modified by Karlovska et al ( 2006 ).…”

Sarco/Endoplasmic Reticulum Calcium ATPase Inhibitors: Beyond Anticancer Perspective

“…Rutin can protect SERCA from the inhibitory effect resulting from treatment of SERCA with hypochloric acid or peroxynitrite. 202,203 Fatty acids are prospective moieties for compound modifications due to their lipophilic properties and related interaction with lipid membranes. Unmodified rutin showed no inhibitory properties at concentrations up to 150 μM; Furthermore, all prepared rutin acyl derivatives continued to inhibit SERCA with similar IC 50 values even in the presence of ionophore ionomycin.…”

“…Another flavonoid examined for SERCA inhibitory activity was rutin and its fatty acid derivatives ( 45 , Figure ). Rutin can protect SERCA from the inhibitory effect resulting from treatment of SERCA with hypochloric acid or peroxynitrite. , Fatty acids are prospective moieties for compound modifications due to their lipophilic properties and related interaction with lipid membranes. Unmodified rutin showed no inhibitory properties at concentrations up to 150 μM; however, rutin stearate, rutin linoleate, and rutin arachidonate exhibited IC 50 values of 35, 25, and 23 μM, respectively.…”