Sphingomyelin phosphodiesterase, also known as acid sphingomyelinase and encoded by the SMPD1 gene, is a lysosomal phosphodiesterase which belongs to the acid sphingomyelinase family (1). SMPD1 catalyzes the hydrolysis of sphingomyelin to ceramide and phosphorylcholine. Ceramide, a bioactive lipid, has emerged as an important signaling molecule involved in a variety of cellular processes such as cell differentiation, apoptosis, and proliferation (2). Activation of SMPD1 occurs by the removal, chemical modification or dimerization of its C-terminal cysteine residue (3). Deficiencies of SMPD1 result in a lysosomal storage disorder referred to as Niemann-Pick disease (4). Recombinant human SMPD1 was expressed without the last three C-terminal residues, and is therefore constitutively active.
Recombinant Human SMPD1 Protein, CF
R&D Systems | Catalog # 5348-PD
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Key Product Details
- R&D Systems CHO-derived Recombinant Human SMPD1 Protein (5348-PD)
- 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
CHO
Accession Number
Applications
Enzyme Activity
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Product Specifications
Source
Chinese Hamster Ovary cell line, CHO-derived human SMPD1 protein
His62-Pro628, with a C-terminal 10-His tag
His62-Pro628, with a C-terminal 10-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
His62
Predicted Molecular Mass
66 kDa
SDS-PAGE
71, 77, and 85 kDa, reducing conditions
Activity
Measured by its ability to cleave 2-N-Hexadecanoylamino-4-nitrophenylphosphorylcholine (HNPPC).
The specific activity is >1,700 pmol/min/μg, as measured under the described conditions.
The specific activity is >1,700 pmol/min/μg, as measured under the described conditions.
Reviewed Applications
Read 1 review rated 5 using 5348-PD in the following applications:
Formulation, Preparation, and Storage
5348-PD
| Formulation | Supplied as a 0.2 μm filtered solution in Tris and NaCl. |
| 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: SMPD1
References
- Schuchman, E.H. et al. (1991) J. Biol. Chem. 266:8531.
- Melendez, A.J. et al. (2008) Biochim. Biophys. Acta 1784:66.
- Qiu, H. et al. (2003) J. Biol. Chem. 278:32744.
- Smith, E.L. and Schuchman, E.H. (2008) FASEB J. 22:3419.
Long Name
Sphingomyelin Phosphodiesterase 1, Acid Lysosomal
Alternate Names
aSMase, NPD
Gene Symbol
SMPD1
UniProt
Additional SMPD1 Products
Product Documents for Recombinant Human SMPD1 Protein, CF
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Recombinant Human SMPD1 Protein, CF
For research use only
Related Research Areas
Citations for Recombinant Human SMPD1 Protein, CF
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Protocols
View specific protocols for Recombinant Human SMPD1 Protein, CF (5348-PD):
Materials
- Assay Buffer: 50 mM MES, 0.5 µM ZnCl2, pH 6.5
- Developing Buffer: 0.2 M NaOH
- Recombinant Human SMPD1 (rhSMPD1) (Catalog # 5348-PD)
- Substrate: 2-N-Hexadecanoyl-4-nitrophenylphosphorylcholine (HNPPC) (Gojira Fine Chemicals, Catalog # HN1004), 50 mM stock solution in methanol. Note: Incubate at 37 °C for 5 minutes and vortex to dissolve.
- 96-well Clear Plate (Catalog # DY990)
- Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
- Dilute rhSMPD1 to 2 µg/mL in Assay Buffer and incubate at 37 °C for 10 minutes.
- Dilute Substrate to 1 mM in Assay Buffer and incubate at 37 °C for 10 minutes.
- Combine 50 µL of rhSMPD1 and 50 µL of Substrate in a plate. Create Substrate Blanks by using Assay Buffer in place of rhSMPD1.
- Cover plate and incubate at room temperature for 20 minutes in the dark.
- Stop the reaction by adding 100 µL of Developing Buffer to each well.
- Read (top read) absorbance in endpoint mode at 410 nm.
- 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 p-Nitrophenol (Sigma-Aldrich, Catalog # 241326).
Per Well:
- rhSMPD1: 0.10 µg
- Substrate: 250 µM