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MUTLU, ÖZAL

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MUTLU

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ÖZAL

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Now showing 1 - 4 of 4
  • Publication
    Biochemical and in silico Characterization of Recombinant L-Lactate Dehydrogenase of Theileria annulata
    (HUMANA PRESS INC, 2016) MUTLU, ÖZAL; Nural, Belma; Erdemir, Aysegul; Mutlu, Ozal; Yakarsonmez, Sinem; Danis, Ozkan; Topuzogullari, Murat; Turgut-Balik, Dilek
    Theileria annulata is a parasite that causes theileriosis in cattle. Reports about drug resistance made essential to develop new drug. LDH of Theileria schizonts is the vital enzyme for its anaerobic metabolism. TaLDH gene was first cloned into pGEM-T cloning vector with two introns in our previous study. Here we report cloning of TaLDH without introns into pLATE 31 vector in E. coli BL21(DE3). Protein was in an inactive form. Two mutations were fixed to express the active protein. Protein was purified by affinity chromatography and evaluated by SDS-PAGE and size exclusion chromatography. Optimum pH of enzyme was performed in pH 7.5, and enzyme was stabilized at 20-40 A degrees C. Enzyme kinetics of recombinant TaLDH were found to be in the direction of pyruvate to lactate K (m) 0.1324 and K (i) 4.295 mM, k (cat) , 44.55/s and k (cat) /K (m) , 3.3693 x 10(5)/M/s. 3D structure of TaLDH was predicted, and possible drug binding sites were determined by homology modelling.
  • Publication
    Discovery and evaluation of inhibitory activity and mechanism of arylcoumarin derivatives on Theileria annulata enolase by in vitro and molecular docking studies
    (SPRINGER, 2020) MUTLU, ÖZAL; Yakarsonmez, Sinem; Danis, Ozkan; Mutlu, Ozal; Topuzogullari, Murat; Sariyer, Emrah; Yuce-Dursun, Basak; Turgut-Balik, Dilek
    In this study, the inhibition potential of 3- and 4-arylcoumarin derivatives on Theileria annulata enolase (TaENO) was assessed for the first time in the literature. Firstly, protein stabilization analyses of TaENO were performed and it was found that the enzyme remains stable with the addition of 6 M ethylene glycol at + 4 degrees C. Inhibitor screening analyses were carried out using 25 coumarin derivatives on highly purified TaENO (> 95%), and four coumarin derivatives [4-(3,4-dimethoxyphenyl)-6,7-dihydroxy-2H-chromen-2-one (C8); 4-(3,4-dihydroxyphenyl)-7,8 dihydroxy-2H-chromen-2-one (C9); 4-(3,4-dihydroxyphenyl)-6,7-dihydroxy-2H-chromen-2 one (C21); and 3-(3,4-dihydroxyphenyl)-7,8-dihydroxy-2H-chromen-2-one (C23)] showed the highest inhibitory effects with the IC50 values of 10.450, 13.170, 8.871 and 10.863 mu M, respectively. The kinetic results indicated that these compounds inhibited the enzyme by uncompetitive inhibition. In addition, the successful binding of the most potent inhibitor (C21) into TaENO was confirmed by using MALDI-TOF mass spectrophotometry. Molecular docking analyses have predicted that C8 and C21 coumarin derivatives which showed high inhibitory effects on TaENO were interacted with high affinity to the potential regions out of the active site. Taken together, these coumarin derivatives (C8, C9, C21 and C23) are first known potent, nonsubstrate, uncompetitive inhibitors of TaENO and these results will facilitate further in vitro and in vivo analysis toward structure-based drug design studies. [GRAPHICS] .
  • Publication
    Comprehensive structural analysis of the open and closed conformations of Theileria annulata enolase by molecular modelling and docking
    (ELSEVIER SCI LTD, 2016) MUTLU, ÖZAL; Mutlu, Ozal; Yakarsonmez, Sinem; Sariyer, Emrah; Danis, Ozkan; Yuce-Dursun, Basalt; Topuzogullari, Murat; Akbulut, Ekrem; Turgut-Balik, Dilek
    Theileria annulata is an apicomplexan parasite which is responsible for tropical theileriosis in cattle. Due to resistance of T. annulata against commonly used antitheilerial drug, new drug candidates should be identified urgently. Enolase might be a druggable protein candidate which has an important role in glycolysis, and could also be related to several cellular functions as a moonlight protein. In this study; we have described three-dimensional models of open and closed conformations of T. annulata enolase by homology modeling method for the first time with the comprehensive domain, active site and docking analyses. Our results show that the enolase has similar folding patterns within enolase superfamily with conserved catalytic loops and active site residues. We have described specific insertions, possible plasminogen binding sites, electrostatic potential surfaces and positively charged pockets as druggable regions in T. annulate enolase. (C) 2016 Elsevier Ltd. All rights reserved.
  • Publication
    Inhibitory effects of arylcoumarin derivatives on Bacteroides fragilis D-lactate dehydrogenase
    (ELSEVIER SCIENCE BV, 2019) MUTLU, ÖZAL; Ugurel, Erennur; Danis, Ozkan; Mutlu, Ozal; Yuce-Dursun, Basak; Gunduz, Cihan; Turgut-Balik, Dilek
    Bacteroides fragilis is an anaerobic bacterium naturally hosted in the human colon flora. B. fragilis D-lactate dehydrogenase (BfD-LDH) is an important enzyme which catalyzes the conversion of D-lactate to pyruvate and regulates anaerobic glycolysis. In this study BfD-LDH has been targeted for structure based drug design. B. fragilis D-lactate dehydrogenase has been expressed, purified and inhibitory activities of 25 coumarin derivatives previously synthetize for their antioxidant activity were evaluated. Among the 25 coumarin derivatives, compound 6a, 51, and 6b exhibited the highest inhibitory activity with IC50 values of 0,47 mu M, 0,57 mu M ye 0,057 mu M, respectively. The results indicate that the mechanism by which 6a, 51 and 6b coumarin derivatives inhibit BfD-LDH by reversible non-competitive inhibition. Docking experiments were carried out to further explain the results and compare the theoretical and experimental affinity of these compounds to the BfD-LDH protein. According to docking results, all coumarins bind to the site occupied by pyruvate and the nicotinamide ring of NADH. (C) 2019 Elsevier B.V. All rights reserved.