A cytosolic glutathione s-transferase, GST-theta from freshwater prawn Macrobrachium rosenbergii: molecular and biochemical properties.

A cytosolic glutathione s-transferase, GST-theta from freshwater prawn Macrobrachium rosenbergii: molecular and biochemical properties.

Gene (2014-06-01)

Jesu Arockiaraj, Annie J Gnanam, Rajesh Palanisamy, Prasanth Bhatt, Venkatesh Kumaresan, Mukesh Kumar Chaurasia, Mukesh Pasupuleti, Harikrishnan Ramaswamy, Abirami Arasu, Akila Sathyamoorthi

PMID24879918

RESUMEN

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 pH7.5 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.

MATERIALES

Número de producto

Marca

Descripción del producto

D2650

Sigma-Aldrich

Dimetilsulfóxido, Hybri-Max^™, sterile-filtered, BioReagent, suitable for hybridoma, ≥99.7%

472301

Sigma-Aldrich

Dimetilsulfóxido, ACS reagent, ≥99.9%

D8418

Sigma-Aldrich

Dimetilsulfóxido, Molecular Biology

34869

Sigma-Aldrich

Dimetilsulfóxido, suitable for HPLC, ≥99.7%

D2438

Sigma-Aldrich

Dimetilsulfóxido, sterile-filtered, BioPerformance Certified, meets EP, USP testing specifications, suitable for hybridoma

D5879

Sigma-Aldrich

Dimetilsulfóxido, ReagentPlus^®, ≥99.5%

D4540

Sigma-Aldrich

Dimetilsulfóxido, ≥99.5% (GC), suitable for plant cell culture

41639

Sigma-Aldrich

Dimetilsulfóxido, BioUltra, Molecular Biology, ≥99.5% (GC)

D1435

Sigma-Aldrich

Dimetilsulfóxido, meets EP testing specifications, meets USP testing specifications

D9170

Sigma-Aldrich

Dimetilsulfóxido, PCR Reagent

391352

Sigma-Aldrich

Lead, powder, −100 mesh, 99.95% trace metals basis

414891

Sigma-Aldrich

Cadmium, granular, 30-80 mesh, ≥99%

G6511

Sigma-Aldrich

Glutathione S-Transferase from equine liver, lyophilized powder, ≥25 units/mg protein

265365

Sigma-Aldrich

Cadmium, powder, −100 mesh, 99.5% trace metals basis

69899

Sigma-Aldrich

Monochlorobimane, suitable for fluorescence, ≥70.0% (HPCE)

94563

Supelco

Dimetilsulfóxido, analytical standard

265330

Sigma-Aldrich

Cadmium, granular, ≥99%, 5-20 mesh

01934

Supelco

Dimetilsulfóxido, for inorganic trace analysis, ≥99.99995% (metals basis)

1211006

USP

Dimetilsulfóxido, United States Pharmacopeia (USP) Reference Standard

396117

Sigma-Aldrich

Lead, shot, <2 mm, 99.9% trace metals basis

GF50662773

Lead, foil, 300x300mm, thickness 0.25mm, as rolled, 99.95%

209708

Sigma-Aldrich

Lead, powder, −325 mesh, ≥99% trace metals basis

695912

Sigma-Aldrich

Lead, shot, 1-3 mm, 99.995% trace metals basis

202886

Sigma-Aldrich

Cadmium, shot, 3 mm, 99.999% trace metals basis

GF64286602

Cadmium, foil, 100x100mm, thickness 1.0mm, as rolled, 99.99%

D2050000

Dimetilsulfóxido, European Pharmacopoeia (EP) Reference Standard

GF05266672

Lead, rod, 100mm, diameter 19.0mm, 99.95%

74875

Sigma-Aldrich

8-Octanoyloxypyrene-1,3,6-trisulfonic acid trisodium salt, suitable for fluorescence, ≥90% (HPCE)

GF22298777

Lead, foil, 150x150mm, thickness 0.1mm, as rolled, 99.95%

GF45457960

Cadmium, foil, 10mm disks, thickness 1.0mm, as rolled, 100%