Dopa Decarboxylase (DDC), also known as aromatic amino acid decarboxylase, is a group II decarboxylase (1). The enzyme catalyzes the decarboxylation of aromatic L-amino acids to produce the corresponding amines. DDC produces the neurotransmitters dopamine and serotonin from L-Dopa and L-5-hydroxytryptophan, respectively. The inhibition of DDC could be used for the treatment of schizophrenia and Parkinson's disease (2).
Recombinant Human Dopa Decarboxylase/DDC Protein, CF
R&D Systems | Catalog # 3564-DC
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Key Product Details
- R&D Systems Sf 21 (baculovirus)-derived Recombinant Human Dopa Decarboxylase/DDC Protein (3564-DC)
- Quality control testing to verify active proteins with lot specific assays by in-house scientists
- All R&D Systems proteins are covered with a 100% guarantee
Source
Sf 21 (baculovirus)
Accession Number
Applications
Enzyme Activity
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Product Specifications
Source
Spodoptera frugiperda, Sf 21 (baculovirus)-derived human Dopa Decarboxylase/DDC protein
Met1-Glu480, with a C-terminal 6-His tag
Met1-Glu480, with a C-terminal 6-His tag
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
N-terminal Sequence Analysis
No result obtained
Predicted Molecular Mass
55 kDa
SDS-PAGE
55 kDa, reducing conditions
Activity
Measured by its ability to convert the substrate 3, 4-dihydroxy L-phenylalanine (L-Dopa) to 3, 4-dihydroxyphenylethylamine (dopamine). The dopamine product is measured by its absorbance at 340 nm after derivatization with trinitrobenzene sulfonic acid. Sherald, F. et al. (1973) Anal. Biochem. 56:300.
The specific activity is >1500 pmol/min/µg, as measured under the described conditions.
The specific activity is >1500 pmol/min/µg, as measured under the described conditions.
Formulation, Preparation, and Storage
3564-DC
| Formulation | Supplied as a 0.2 μm filtered solution in MES, NaCl and DTT. |
| Shipping | The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
|
Background: Dopa Decarboxylase/DDC
References
- Sandmeier, E. et al. (1994) Eur. J. Biochem. 221:997.
- Bertoldi, M. et al. (1996) J. Biol. Chem. 271:23954.
Alternate Names
AADC, DDC
Entrez Gene IDs
1644 (Human)
Gene Symbol
DDC
UniProt
Additional Dopa Decarboxylase/DDC Products
Product Documents for Recombinant Human Dopa Decarboxylase/DDC Protein, CF
Certificate of Analysis
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Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Recombinant Human Dopa Decarboxylase/DDC Protein, CF
For research use only
Related Research Areas
Citations for Recombinant Human Dopa Decarboxylase/DDC Protein, CF
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Protocols
View specific protocols for Recombinant Human Dopa Decarboxylase/DDC Protein, CF (3564-DC):
Materials
- Assay Buffer: 50 mM HEPES, 100 mM NaCl, pH 7.2
- Recombinant Human Dopa Decarboxylase/DDC (rhDopa) (Catalog # 3564-DC)
- Pyridoxal 5’-Phosphate hydrate (Sigma, Catalog # P9255)
- 3,4-dihydroxy L-phenylalanine (L-DOPA) (Sigma, Catalog # D9628)
- 2,4,6-Trinitrobenzene sulfonic acid (TNBS), 5% picryl solution (Sigma, Catalog # P2297)
- Benzene (Sigma, Catalog # 319953)
- 96 well clear UV-transparent microplate (Corning, Catalog # 3635)
- Plate Reader (Model: Spectramax Plus by Molecular Devices) or equivalent
- Prepare 5 mM stock solutions of L-DOPA and pyridoxal phosphate in deionized water. Use vigorous vortexing and mixing to get both reagents into solution.
- Dilute rhDopa to 5 ng/µL in Assay Buffer.
- Combine 160 µL of 5 ng/μL rhDopa with 20 µL 5 mM L-DOPA, 4 µL 5 mM pyridoxal phosphate, and 16 µL Assay Buffer. Include a control containing 20 µL 5 mM L-DOPA, 4 µL 5 mM pyridoxal phosphate, and 176 µL Assay Buffer without any rhDopa.
- Incubate for 30 minutes at 37 °C.
- Stop the reaction by heating for 2 minutes at 95 - 100 °C. After heating, cool on ice.
- Add 10 µL of 5% TNBS to each tube and vortex.
- Incubate for 20 minutes at 37 °C. Ensure that all samples undergo derivitization for the same length of time, stagger the addition of TNBS and the benzene extraction (next step) if necessary.
- In a fume hood, add 300 µL of benzene to each reaction in the same order the 5% TNBS was added. Vortex for 30 seconds.
- Centrifuge all reactions at 13,000 rpm in a microcentrifuge for 5 minutes to separate the aqueous and organic solvents.
- Carefully pipet out 200 µL of the benzene phase (the upper layer) of each reaction and load into a UV-transparent microplate. It is recommended that the microplate be kept covered as much as possible to minimize evaporation of the benzene.
- Read at an absorbance of 340 nm in endpoint mode.
- Calculate specific activity:
|
Specific Activity (pmol/min/µg) = |
Adjusted Abs* (OD) x Conversion Factor** (pmol/OD) |
| Incubation time (min) x amount of enzyme (µg) |
*Adjusted for Substrate Blank
**Derived using calibration standard dopamine (3,4-dihydroxyphenylethylamine) (Sigma, Catalog # H8502).
Per Reaction:- rhDopa: 0.800 µg
- L-DOPA: 500 µM
- Pyridoxal Phosphate: 100 µM
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