Identification and characterization of the zebrafish glutathione S‐transferase Pi‐1

Abunnaja, Maryam S.; Kurogi, Katsuhisa; Mohammed, Yasir I.; Sakakibara, Yoichi; Suiko, Masahito; Hassoun, Ezdihar A.; Liu, Ming‐Cheh

doi:10.1002/jbt.21948

Cited by 7 publications

(7 citation statements)

References 51 publications

(69 reference statements)

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“…The Gst superfamily enzymes primarily function as detoxification enzymes, and are highly evolutionarily conserved [25,61]. In the zebrafish, Gst enzymes follow expression patterns similar to those seen in humans; this is true at both transcriptomic and proteomic levels [24,49,62,63]. MCB fluorescence patterns during embryonic development mirror Gst expression and embryonic GSH levels, and, correlate with key developmental events.…”

Section: Discussionmentioning

confidence: 99%

Mapping glutathione utilization in the developing zebrafish (Danio rerio) embryo

et al. 2019

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Glutathione (GSH), the most abundant vertebrate endogenous redox buffer, plays key roles in organogenesis and embryonic development, however, organ-specific GSH utilization during development remains understudied. Monochlorobimane (MCB), a dye conjugated with GSH by glutathione-s-transferase (GST) to form a fluorescent adduct, was used to visualize organ-specific GSH utilization in live developing zebrafish ( Danio rerio ) embryos. Embryos were incubated in 20 μM MCB for 1 h and imaged on an epifluorescence microscope. GSH conjugation with MCB was high during early organogenesis, decreasing as embryos aged. The heart had fluorescence 21-fold above autofluorescence at 24 hpf, dropping to 8.5-fold by 48 hpf; this increased again by 72 hpf to 23.5-fold, and stayed high till 96 hpf (18-fold). The brain had lower fluorescence (10-fold) at 24 and 48 hpf, steadily increasing to 30-fold by 96 hpf. The sensitivity and specificity of MCB staining was then tested with known GSH modulators. A 10-min treatment at 48 hpf with 750 μM tert -butylhydroperoxide, caused organ-specific reductions in staining, with the heart losing 30% fluorescence, and, the brain ventricle losing 47% fluorescence. A 24 h treatment from 24-48 hpf with 100 μM of N-Acetylcysteine (NAC) resulted in significantly increased fluorescence, with the brain ventricle and heart showing 312% and 240% increases respectively, these were abolished upon co-treatment with 5 μM BSO, an inhibitor of the enzyme that utilizes NAC to synthesize GSH. A 60 min 100 μM treatment with ethacrynic acid, a specific GST inhibitor, caused 30% reduction in fluorescence across all measured structures. MCB staining was then applied to test for GSH disruptions caused by the toxicants perfluorooctanesulfonic acid and mono-(2-ethyl-hexyl)phthalate; MCB fluorescence responded in a dose, structure and age-dependent manner. MCB staining is a robust, sensitive method to detect spatiotemporal changes in GSH utilization, and, can be applied to identify sensitive target tissues of toxicants.

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Section: Discussionmentioning

confidence: 99%

Mapping glutathione utilization in the developing zebrafish (Danio rerio) embryo

et al. 2019

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“…Additionally, GSTs can catalyze the combination of glutathione (GSH) and various active electrophiles through affinity attacks and couplings [4]. GSTs have been widely described from animals [5], plants [6], and microorganisms [7]. Recently, a GST with an optimum temperature of 40 °C has been identified in the Antarctic psychrophilic bacteria Pseudoalteromonas sp.…”

Section: Introductionmentioning

confidence: 99%

Cold-Adapted Glutathione S-Transferases from Antarctic Psychrophilic Bacterium Halomonas sp. ANT108: Heterologous Expression, Characterization, and Oxidative Resistance

et al. 2019

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Glutathione S-transferases are one of the most important antioxidant enzymes to protect against oxidative damage induced by reactive oxygen species. In this study, a novel gst gene, designated as hsgst, was derived from Antarctic sea ice bacterium Halomonas sp. ANT108 and expressed in Escherichia coli (E. coli) BL21. The hsgst gene was 603 bp in length and encoded a protein of 200 amino acids. Compared with the mesophilic EcGST, homology modeling indicated HsGST had some structural characteristics of cold-adapted enzymes, such as higher frequency of glycine residues, lower frequency of proline and arginine residues, and reduced electrostatic interactions, which might be in relation to the high catalytic efficiency at low temperature. The recombinant HsGST (rHsGST) was purified to apparent homogeneity with Ni-affinity chromatography and its biochemical properties were investigated. The specific activity of the purified rHsGST was 254.20 nmol/min/mg. The optimum temperature and pH of enzyme were 25 °C and 7.5, respectively. Most importantly, rHsGST retained 41.67% of its maximal activity at 0 °C. 2.0 M NaCl and 0.2% H2O2 had no effect on the enzyme activity. Moreover, rHsGST exhibited its protective effects against oxidative stresses in E. coli cells. Due to its high catalytic efficiency and oxidative resistance at low temperature, rHsGST may be a potential candidate as antioxidant in low temperature health foods.

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“…To date, zebrafish GST studies have focused on 2 levels: GST enzymatic activity ( Best et al , 2002 ; Notch et al , 2011 ; Otte et al , 2017 ; Pavagadhi et al , 2012 ; Wiegand et al , 2000 ) and mRNA expression ( Abunnaja et al , 2017 ; Glisic et al , 2015 , 2016 ; Timme-Laragy et al , 2013 ). GST enzymatic activity was detected within the first 4 h of zebrafish development ( Notch et al , 2011 ; Otte et al , 2017 ; Wiegand et al , 2000 ) as well as in all examined organs of adult zebrafish ( Pavagadhi et al , 2012 ).…”

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confidence: 99%

Glutathione S-Transferase Protein Expression in Different Life Stages of Zebrafish (Danio rerio)

Tierbach

Groh

Schönenberger

et al. 2018

Toxicological Sciences

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Zebrafish is a widely used animal model in biomedical sciences and toxicology. Although evidence for the presence of phases I and II xenobiotic defense mechanisms in zebrafish exists on the transcriptional and enzyme activity level, little is known about the protein expression of xenobiotic metabolizing enzymes. Given the important role of glutathione S-transferases (GSTs) in phase II biotransformation, we analyzed cytosolic GST proteins in zebrafish early life stages and different organs of adult male and female fish, using a targeted proteomics approach. The established multiple reaction monitoring-based assays enable the measurement of the relative abundance of specific GST isoenzymes and GST classes in zebrafish through a combination of proteotypic peptides and peptides shared within the same class. GSTs of the classes alpha, mu, pi and rho are expressed in zebrafish embryo as early as 4 h postfertilization (hpf). The majority of GST enzymes are present at 72 hpf followed by a continuous increase in expression thereafter. In adult zebrafish, GST expression is organ dependent, with most of the GST classes showing the highest expression in the liver. The expression of a wide range of cytosolic GST isoenzymes and classes in zebrafish early life stages and adulthood supports the use of zebrafish as a model organism in chemical-related investigations.

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Identification and characterization of the zebrafish glutathione S‐transferase Pi‐1

Cited by 7 publications

References 51 publications

Mapping glutathione utilization in the developing zebrafish (Danio rerio) embryo

Mapping glutathione utilization in the developing zebrafish (Danio rerio) embryo

Cold-Adapted Glutathione S-Transferases from Antarctic Psychrophilic Bacterium Halomonas sp. ANT108: Heterologous Expression, Characterization, and Oxidative Resistance

Glutathione S-Transferase Protein Expression in Different Life Stages of Zebrafish (Danio rerio)

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