AKR1A1 Protein, Human (His)

The AKR1A1 protein uses NADPH to reduce various carbonyl compounds to alcohols. AKR1A1 Protein, Human (His) is the recombinant human-derived AKR1A1 protein, expressed by E. coli , with N-His labeled tag.

AKR1A1, a versatile enzyme, catalyzes the NADPH-dependent reduction of a broad spectrum of carbonyl-containing compounds to their respective alcohols. Exhibiting enzymatic activity towards endogenous metabolites such as aromatic and aliphatic aldehydes, ketones, monosaccharides, and bile acids, AKR1A1 demonstrates a preference for negatively charged substrates like glucuronate and succinic semialdehyde. Functioning as a detoxifying enzyme, it plays a crucial role in reducing toxic aldehydes, including methylglyoxal and 3-deoxyglucosone, which accumulate under hyperglycemic conditions and exert cytotoxic effects. Additionally, AKR1A1 contributes to the detoxification of lipid-derived aldehydes, such as acrolein. The enzyme is implicated in the activation of procarcinogens, including polycyclic aromatic hydrocarbon trans-dihydrodiols, and participates in the metabolism of various xenobiotics and drugs, such as the anthracyclines doxorubicin (DOX) and daunorubicin (DAUN). Notably, AKR1A1 does not exhibit reductase activity towards retinoids. This diverse substrate specificity underscores its pivotal role in cellular detoxification processes and its potential relevance in therapeutic interventions.

Measured by its ability to Reduction of ketone. The specific activity is 2227.65 pmol/min/μg, as measured under the described conditions.

Materials
AKR1A1 Protein, Human (His) (HY-P76137)
Assay Buffer: 100 mM Sodium Phosphate Buffer, pH 7.0
Substrate: 4-Nitrobenzaldehyde
Standard: NADPH

Procedure
1. Preparation of standard curve: Prepare NADPH standard with assay buffer to concentrations of 2, 1, 0.5, 0.25, 0.1, 0.05, 0 mM. Add 200 μL per well, measure OD₃₄₀, and plot a standard curve with concentration as the abscissa and OD value as the ordinate.
2. Mixture: Prepare a solution containing 0.2 mM NADPH and 1 mM 4-Nitrobenzaldehyde using assay buffer.
3. Dilute Human AKR1A1 protein to 100 μg/mL with assay buffer.
4. Experimental group: 150 μL mixture + 50 μL Human AKR1A1.
Blank group: 200 μL mixture.
5. Measure OD₃₄₀ in kinetic mode for 5 min at 24°C.
6. Calculate the specific activity:

Human

E. coli

N-His

P14550 (M1-Y325)

10327  [NCBI]

Full Length

MAASCVLLHTGQKMPLIGLGTWKSEPGQVKAAVKYALSVGYRHIDCAAIYGNEPEIGEALKEDVGPGKAVPREELFVTSKLWNTKHHPEDVEPALRKTLADLQLEYLDLYLMHWPYAFERGDNPFPKNADGTICYDSTHYKETWKALEALVAKGLVQALGLSNFNSRQIDDILSVASVRPAVLQVECHPYLAQNELIAHCQARGLEVTAYSPLGSSDRAWRDPDEPVLLEEPVVLALAEKYGRSPAQILLRWQVQRKVICIPKSITPSRILQNIKVFDFTFSPEEMKQLNALNKNWRYIVPMLTVDGKRVPRDAGHPLYPFNDPY

Approximately 39 kDa.

≥ 90%, as determined by reducing SDS-PAGE.

Lyophilized powder.

Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.5, 5% trehalose, 5% mannitol and 0.01% Tween 80.

<1 EU/μg, determined by LAL method.

It is not recommended to reconstitute to a concentration less than 100 μg/mL in ddH₂O.

Stored at -20°C for 2 years from date of receipt. After reconstitution, it is stable at 4°C for 1 week or -20°C for longer (with carrier protein). It is recommended to freeze aliquots at -20°C or -80°C for extended storage.

Room temperature in continental US; may vary elsewhere.