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A cytosolic glutathione s-transferase,GST-theta from freshwater prawn Macrobrachium rosenbergii: molecular and biochemical properties
Authors:Jesu Arockiaraj  Annie J Gnanam  Rajesh Palanisamy  Prasanth Bhatt  Venkatesh Kumaresan  Mukesh Kumar Chaurasia  Mukesh Pasupuleti  Harikrishnan Ramaswamy  Abirami Arasu  Akila Sathyamoorthi
Institution:1. Division of Fisheries Biotechnology & Molecular Biology, Department of Biotechnology, Faculty of Science and Humanities, SRM University, Kattankulathur 603 203, Chennai, Tamil Nadu, India;2. Institute for Cellular and Molecular Biology, The University of Texas at Austin, 1 University Station A4800, Austin, TX 78712, USA;3. Lab PCN 206, Microbiology Division, CSIR-Central Drug Research Institute, B.S. 10/1, Sector 10, Jankipuram Extension, Sitapur Road, Lucknow 226031, Uttar Pradesh, India;4. PG and Research Department of Biotechnology, Bharath College of Science and Management, Thanjavur 613 005, Tamil Nadu, India;5. Department of Microbiology, SRM Arts & Science College, Kattankulathur 603 203, Chennai, India;6. Department of Biotechnology, SRM Arts & Science College, Kattankulathur 603 203, Chennai, India
Abstract:Glutathione S-transferases play an important role in cellular detoxification and may have evolved to protect cells against reactive oxygen metabolites. In this study, we report the molecular characterization of glutathione s-transferase-theta (GST-θ) from freshwater prawn Macrobrachium rosenbergii. A full length cDNA of GSTT (1417 base pairs) was isolated and characterized bioinformatically. Exposure to virus (white spot syndrome baculovirus or M. rosenbergii nodovirus), bacteria (Aeromonas hydrophila or Vibrio harveyi) or heavy metals (cadmium or lead) significantly increased the expression of GSTT (P < 0.05) in hepatopancreas. Recombinant GST-θ with monochlorobimane substrate had an optimum activity at pH 7.5 and 35 °C. Furthermore recombinant GST-θ activity was abolished by the denaturants triton X-100, Gua-HCl, Gua-thiocyanate, SDS and urea in a dose-dependent manner. Overall, the results suggest a potential role for M. rosenbergii GST-θ in detoxification and possibly conferring immune protection.
Keywords:Mr  Macrobrachium rosenbergii  GST-θ  glutathione s-transferase-theta  MrGST-θ  Macrobrachium rosenbergii glutathione s-transferase-theta  GSH  glutathione  MAPEG  membrane-associated proteins in eicosanoid and glutathione metabolism  WSBV  white spot syndrome baculovirus  MrNV  Macrobrachium rosenbergii nodovirus  MCB  monochlorobimane  Cd  cadmium  Pb  lead  ORF  open reading frame  UTR  untranslated region  qRT-PCR  quantitative real time polymerase chain reaction  IPTG  isopropyl-β-thiogalactopyranoside
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