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Encompassing Amino Acids: 147-end
Applications: Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Assay Conditions: ALK2 (R206H) activity was measured by using the casein protein substrate diluted in distilled water to a working concentration of 1 mg/ml, in the ADP Glo™ Kinase Assay kit (Promega #V9101). Reaction was initiated by mixing increasing amounts of ALK2 (R206H) with 25 µM ATP in 40 mM Tris-HCl, pH 7.4, 20 mM MgCl2, 0.1 mg/ml BSA prepared with 50 µM DTT and substrate at a final concentration of 200 µg/ml.After a 40-minute incubation at room temperature, the reaction was terminated by addition of ADP-Glo™ Reagent, followed by a subsequent 40-minute incubation at room temperature. Kinase Detection Reagent was added, and the reaction was incubated for another 30 minutes at ambient temperature. Detection of luminescence was measured using the Luminescence Module Protocol on GloMax®-Multi Microplate Multimode Reader. The Specific Activity was calculated as follows: (Corrected activity, RLU) / [(Specific activity from ADP in RLU/pmol) * (Reaction time in min) *(Enzyme amount in µg or mg)]. Corrected RLU was calculated by subtracting the blank value from all the values. The blank was determined from a "no enzyme" sample by replacing the enzyme solution with an equal volume of Dilution Buffer IX (1x).
Background: ALK2 (activin receptor-like kinase 2), also known as ACVR1 or activin A receptor type 1, is a bone morphogenic protein receptor involved in BMP (bone morphogenic protein) signal transduction. ALK2 forms complexes with BMPs, which then recruit proteins from the SMAD family (Mothers against decapentaplegic homolog). It participates in the development and regulation of the skeletal system, heart, brain and reproductive system. ALK2 dysfunction can lead to fibrodysplasia ossificans progressive (FOP), where the BMP/SMAD pathway is hyper-activated and mesenchymal stem cells differentiate along the osteogenic pathway and transform into bone all over the body. Mutations in ALK2 were also found in cancer, such as diffuse intrinsic pontine glioma (DIPG) and child brain cancer. ALK2 inhibitors have been studied in pre-clinical models of FOP and DIPG and showed great promise. Further studies into ALK2 will deepen our understanding of its functions, find new inhibitors and new therapeutic avenues for patients with ALK-linked cancer.
Description: Recombinant mouse ALK2 (R206H) (activin receptor-like kinase-2), encompassing amino acids 147-end with an R206H mutation. This construct contains an N-terminal GST-tag. The recombinant protein was affinity purified and is active.
Format: Aqueous buffer solution
Formulation: 50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 10 mM Glutathione, 0.1 mM EDTA, 0.25 mM DTT, 25% glycerol
Purity: ≥90% (Purity calculation does not include co-purifying Glutathione-binding proteins.)
Storage Stability: At least 6 months at -80°C.
Tags: N-terminal GST-tag
Uniprot: P37172-1
Warnings: Avoid freeze/thaw cycles
Biosafety Level: Not applicable (BSL-1)
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