Vaccinia Virus VH1-related Phosphatase (VHR), also known as Dual-Specificity Phosphatase 3 (DUSP3), removes inorganic phosphate groups covalently attached to tyrosine, serine and threonine residues in proteins (1). It belongs to a family of phosphatases that selectively dephosphorylate MAP kinases. VHR has been shown to dephosphorylate cellular ERK1, ERK2 and JNK, but not p38 (2, 3). Phosphorylation of VHR at tyrosine 138 by ZAP-70 enhances inhibition of ERK2, suggesting a role for VHR in modulating T-cell receptor responses (3). The enzymatic mechanism for VHR has been studied in detail (4) and its X-ray crystal structure has been characterized (5). Its well-defined biochemistry has made it useful in screening assays for compounds that inhibit phosphatases (6).
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Key Product Details
- R&D Systems E. coli-derived Recombinant Human VHR Protein (1654-VH)
- 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
E. coli
Accession Number
Applications
Enzyme Activity
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Product Specifications
Source
E. coli-derived human VHR protein
Ser2-Pro185, with an N-terminal Met and 6-His tag
Ser2-Pro185, with an N-terminal Met and 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
Met
Predicted Molecular Mass
21 kDa
SDS-PAGE
23 kDa, reducing conditions
Activity
Measured by its ability to cleave a substrate, p-Nitrophenyl phosphate (pNPP).
The specific activity is >100 pmol/min/μg, as measured under the described conditions.
The specific activity is >100 pmol/min/μg, as measured under the described conditions.
Formulation, Preparation, and Storage
1654-VH
| Formulation | Supplied as a 0.2 μm filtered solution in HEPES, NaCl, DTT and Glycerol. |
| 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: VHR
References
- Ishibashi, T. et al. (1992) Proc. Natl. Acad. Sci. USA 89:12170.
- Todd, J.L. et al. (1999) J. Biol. Chem. 274:13271.
- Alonso, A. et al. (2003) Nat. Immunol. 4:44.
- Zhang, Z.-Y. et al. (1995) Biochemistry 34:16088.
- Yuvaniyama, J. et al. (1996) Science 272:1328.
- Ueda, K. et al. (2002) FEBS Lett. 525:48.
Long Name
Vaccinia Virus VH1-related Phosphatase
Alternate Names
DUSP3
Entrez Gene IDs
1845 (Human)
Gene Symbol
DUSP3
UniProt
Additional VHR Products
Product Documents for Recombinant Human VHR 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 VHR Protein, CF
For research use only
Related Research Areas
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Protocols
View specific protocols for Recombinant Human VHR Protein, CF (1654-VH):
Materials
- Assay Buffer: 50 mM HEPES, 10 mM DTT, pH 7.5
- Recombinant Human VHR (rhVHR) (Catalog # 1654-VH)
- Substrate: p-Nitrophenyl phosphate (pNPP), 10 mM stock in deionized water
- Sodium Hydroxide (NaOH), 0.2 M stock in deionized water
- Clear 96-well Plate (Catalog # DY990)
- Plate Reader with absorbance read capability
- Dilute rhVHR to 20 µg/mL in Assay Buffer.
- Dilute Substrate to 6 mM in Assay Buffer.
- Prepare reaction mixtures by combining equal volumes of dilute rhVHR and dilute Substrate in microtubes. Include an Enzyme Control by combining dilute rhVHR with twice the volume of 0.2 M NaOH, mix briefly; then add a volume of dilute Substrate equal the volume of dilute rhVHR. The Enzyme Control will have 2x the volume of the reaction mixture.
- Incubate Reactions and Enzyme Control at 37 °C for 4 hours.
- Load 100 µL of Reactions into a plate and stop the reactions by adding 100 µL 0.2 M NaOH.
- Load 200 µL of Enzyme Controls into plate.
- Read plate at 410 nm (absorbance) 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 Enzyme Controls
**Derived using calibration standard p-Nitrophenol
Per Well:
- rhVHR: 1 µg
- pNPP: 1.5 mM
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