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Cat. No.: SMTGR-250 (for 250μg)

Cat. No.: SMTGR-1k (for 1mg)

Description

Thioredoxin Glutathione Reductase (SmTGR), namely Schistosoma mansoni Thioredoxin Glutathione Reductase (SmTGR). Schistosoma mansoni lacks thioredoxin reductase and glutathione reductase. Thioredoxin glutathione reductase (TGR) possesses both thioredoxin reductase and glutathione reductase activities. This enzyme can reduce oxidized glutathione (GSSG) to reduced glutathione (GSH) in an NADPH-dependent manner; it is also capable of reducing thioredoxin (Trx) and 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB).

The catalytic reaction mediated by SmTGR is described as follows: Using NADPH as the electron donor, SmTGR reduces GSSG, thioredoxin (Trx) and DTNB to produce GSH, Trx-(SH)₂ and TNB, respectively, with the concomitant formation of NADP⁺. The activity of SmTGR can be determined via the reduction of DTNB catalyzed by SmTGR.

SmTGR is a unique selenoprotein featuring a GCUG motif at its carboxyl terminus. The penultimate amino acid residue is selenocysteine (Sec, U), known as the 21st naturally occurring amino acid. Structurally highly homologous to cysteine, selenocysteine is a highly reactive amino acid. Compared with cysteine, Sec exhibits stronger nucleophilicity and a lower redox potential, and it is indispensable for the catalytic activity of SmTGR .
As a crucial enzyme in the antioxidant defense system, SmTGR reduces GSSG to GSH. GSH lowers intracellular oxidative levels and thereby alleviates oxidative stress. In addition, it maintains intracellular redox balance and modulates signal transduction by reducing thioredoxin, further governing cell proliferation, differentiation, cell death and other vital physiological processes. Silencing the TGR gene via RNA interference leads to rapid parasite death, making TGR a promising drug target for schistosomiasis treatment.

Applications

Catalyze the reduction of GSSG to GSH; reduce DTNB to TNB for NADPH quantification; screen inhibitors of thioredoxin glutathione reductase; support drug development for schistosomiasis and other trematodiases.

Source

Recombinant protein expressed in Escherichia coli using the TGR gene from Schistosoma mansoni.

Activity Definition

One unit is defined as the amount of enzyme required to reduce 1 μmol DTNB per minute in 0.5 mL standard DTNB assay system containing 2.5 mM DTNB and 0.3 mM NADPH in TE Buffer (50 mM Tris-HCl, 2 mM EDTA, pH 7.5 at 25 °C).

Specific Activity

0.1 U/mg

Purity

Free of DNA endonuclease, exonuclease, RNase and phosphatase activities.

Enzyme Storage Buffer

50 mM Tris-HCl (pH 7.5 at 25 °C), 2 mM EDTA, 50% Glycerol.

Storage Conditions​

Store at -20 °C; stable for at least one year.

Precautions

• Assays with this product involve redox reactions, so any oxidizing or reducing agents will interfere with the measurement. If reducing agents in samples such as DTT and β-mercaptoethanol cannot be avoided, their total concentration should be kept below 0.1 mM. DTT at 0.15 mM can inhibit 40% of the enzyme activity. In addition, sodium sulfate, ammonium sulfate, ferricyanide and other substances may interfere with enzyme activity; avoid their presence as much as possible.
• NADPH is relatively unstable. Store it at low temperature or prepare fresh solutions before use.
• Reaction temperature must be strictly controlled; otherwise, significant measurement errors will occur.
• This product is intended solely for scientific research by professionals. It shall not be used for clinical diagnosis or therapy, food or pharmaceutical applications, and must not be stored in residential premises.
• For personal safety and health, wear a lab coat and disposable gloves during handling.

Only for research and not intended for treatment of humans or animals

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