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The Interaction of Schistosoma Japonicum Glutathione Transferase with Cibacron Blue 3GA and its Fragments

Author(s):

Michalis Platis, Dimitrios Vlachakis, Ahmed Ibrahim Foudah, Magdy Mohamed Muharram, Mohamed Hamed Alqarni, Anastassios C. Papageorgiou and Nikolaos E. Labrou*   Pages 1 - 12 ( 12 )

Abstract:


Background: The 26kDa glutathione transferase (GST, EC 2.5.1.18) from Schistosoma japonicum (SjGST) is recognized as the major detoxification enzyme of S. japonicum, a pathogenic helminth causing schistosomiasis.

Objective: In the present study, the interaction of the chlorotriazine dye Cibacron blue 3GA (CB3GA) and its structural analogues with SjGST was investigated. The work aimed to shine light on the non-substrate ligand-binding properties of the enzyme.

Methods: Kinetic inhibition analysis, affinity labelling experiments and molecular modelling studies were employed.

Results: The results showed that CB3GA is a potent inhibitor (IC50 0.057 ± 0.003μM) towards SjGST. The enzyme was specifically and irreversibly inactivated by the dichlorotriazine-analogue of CB3GA (IC50 0.190 ± 0.024 μM), following a biphasic pseudo-first-order saturation kinetics with approximately 1 mol of inhibitor per mol of dimeric enzyme being incorporated. All other monochlorotriazine analogues behave as reversible inhibitors with lower inhibition potency (IC50 5.2-82.3 μM). Kinetic inhibition studies together with molecular modelling and molecular dynamics simulations established that the CB3GA binding site overlaps both the G- and H-sites. Both hydrophobic/polar interactions as well as steric effects have decisive roles in determining the inhibitory strength of CB3GA and its analogues.

Conclusion: The results of the present study might be useful in future drug design and development efforts towards SjGST.

Keywords:

Affinity labelling, Cibacron Blue 3GA, Schistosoma japonicum, Schistosomiasis, Structure-guided drug design, Triazine dyes.

Affiliation:

Laboratory of Enzyme Technology, Department of Biotechnology, School of Applied Biology and Biotechnology, Agricultural University of Athens, 75 Iera Odos Street, GR-11855-Athens, Laboratory of Genetics, Department of Biotechnology, School of Applied Biology and Biotechnology, Agricultural University of Athens, 75 Iera Odos Street, GR11855-Athens, Department of Pharmacognosy, College of Pharmacy, Prince Sattam Bin Abdulaziz University, 11942 Al Kharj, Department of Pharmaceutics, College of Pharmacy, Prince Sattam Bin Abdulaziz University, 11942, Alkharj, Department of Pharmacognosy, College of Pharmacy, Prince Sattam Bin Abdulaziz University, 11942, Alkharj, Turku Bioscience Centre, University of Turku and Åbo Akademi University, Turku 20521, Laboratory of Enzyme Technology, Department of Biotechnology, School of Applied Biology and Biotechnology, Agricultural University of Athens, 75 Iera Odos Street, GR-11855-Athens



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