SMPD1 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 into 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 (Cys 627) (3). Deficiencies of SMPD1 result in a lysosomal storage disorder referred to as Niemann-Pick disease (4). rmSMPD1 was expressed without its C-terminal cysteine residue, and is therefore constitutively active.
Recombinant Mouse SMPD1 Protein, CF
R&D Systems | Catalog # 5680-PD
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Key Product Details
- R&D Systems CHO-derived Recombinant Mouse SMPD1 Protein (5680-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
Structure / Form
Monomer
Applications
Enzyme Activity
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Product Specifications
Source
Chinese Hamster Ovary cell line, CHO-derived mouse SMPD1 protein
Tyr58-Leu626, with a C-terminal 10-His tag
Tyr58-Leu626, 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
Tyr58
Predicted Molecular Mass
65 kDa
SDS-PAGE
70-80 kDa, reducing conditions
Activity
Measured by its ability to cleave 2-N-Hexadecanoylamino-4-nitrophenylphosphorylcholine (HNPPC).
The specific activity is >1000 pmol/min/μg, as measured under the described conditions.
The specific activity is >1000 pmol/min/μg, as measured under the described conditions.
Formulation, Preparation, and Storage
5680-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.
- Qui, H. et al. (2003) J. Biol. Chem. 278:32744.
- Smith, E.L. and E.H. Schuchman (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 Mouse 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 Mouse SMPD1 Protein, CF
For research use only
Related Research Areas
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Protocols
View specific protocols for Recombinant Mouse SMPD1 Protein, CF (5680-PD):
Materials
- Assay Buffer: 50 mM MES, 0.5 µM ZnCl2, pH 6.5
- Developing Buffer: 0.2 M NaOH
- Recombinant Mouse SMPD-1 (rmSMPD-1) (Catalog # 5680-PD)
- Substrate: 2-N-Hexadecanoylamino-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 rmSMPD-1 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 rmSMPD-1 and 50 µL of Substrate in a plate. Create Substrate Blanks by using Assay Buffer in place of rmSMPD-1.
- 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:
- rmSMPD-1: 0.10 µg
- Substrate: 250 µM