Functional analyses of dipeptide and pentapeptide insertions on Theileria annulata enolase by site-directed mutagenesis and in silico approaches.

Autor: Yakarsonmez S; Istanbul Yeni Yuzyil University, Faculty of Pharmacy, Department of Pharmaceutical Biotechnology, 34010 Istanbul, Turkey., Cengiz EC; Institute for Cellular and Molecular Biology, The University of Texas at Austin, Austin, TX 78712, USA., Mutlu O; Marmara University, Faculty of Arts and Sciences, Department of Biology, 34722 Istanbul, Turkey., Turgut-Balik D; Yıldız Technical University, Faculty of Chemical and Metallurgical Engineering, Department of Bioengineering, 34210 Istanbul, Turkey. Electronic address: dilekbalik@gmail.com.
Jazyk: angličtina
Zdroj: Biochimica et biophysica acta. Proteins and proteomics [Biochim Biophys Acta Proteins Proteom] 2019 Jul - Aug; Vol. 1867 (7-8), pp. 732-739. Date of Electronic Publication: 2019 May 16.
DOI: 10.1016/j.bbapap.2019.05.006
Abstrakt: Theileria annulata enolase (TaENO) could be assessed as a druggable target for tropical theileriosis treatment. The parasite enzyme plays an important role in many cellular functions and carries some structural differences like dipeptide ( 262 EK 263 ) and pentapeptide ( 103 EWGYC 107 ) insertions from the host enzyme, Bos taurus enolase. In this study, the functional effects of these insertions on TaENO activity were analyzed by in vitro site-directed mutagenesis and in silico molecular docking analyses for the first time in the literature. In vitro results showed that, although the deletion of the pentapeptide insertion (TaENOΔEWGYC) reduced the enzyme activity slightly, the removal of the dipeptide insertion (TaENOΔEK) halted it. Also, molecular docking results revealed that the deletion of these insertions affected the substrate binding affinity of the mutant enzymes. The active site of TaENOΔEK exhibited a small decrease of substrate binding affinity compared to the active site of TaENOΔEWGYC relative to the wild type TaENO. Although we conclude that both regions could be evaluated as possible drug-binding sites to inhibit TaENO in further studies, these results indicate that the dipeptide insertion could be a more promising drug binding site than the pentapeptide insertion.
(Copyright © 2019 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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