The Kinase Enzyme Systems include a recombinant kinase enzyme, a substrate appropriate for the enzyme, a reaction buffer and supplemental reagents as needed. The TMP3-ROS1 Kinase Enzyme System contains:
- TMP3-ROS1 Kinase, 10μg [Human, recombinant; amino acids 1–258 (TPM3) and 1881–end (ROS1)]. MW: ~108kDa
- IGF1Rtide Substrate (1mg/ml), (KKKSPGEYVNIEFG), derived from human IRS-1 protein residues 891–902
- 5X Reaction Buffer, 0.1M DTT
Recombinant human fusion protein TPM3 (amino acids 1–258)-ROS1 (amino acids 1881–end) was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. TPM3 is a member of the tropomyosin family of actin-binding proteins which are dimers of coiled-coil proteins that provide stability to actin filaments and regulate access of other actin-binding proteins. Mutations in TPM3 gene result in autosomal dominant nemaline myopathy and other muscle disorders (1). TPM3 gene locus is involved in translocations with other loci including c-ros oncogene 1 (ROS1) which result in the formation of the TPM3-ROS1 fusion protein that act as an oncogene. TPM3-ROS1 fusion gene product has been detected in lung adenocarcinoma is higher in lung adenocarcinoma with wild-type EGFR (2).
Application Note
TMP3 NCBI Database Entry
ROS1 NCBI Database Entry
The TMP3-ROS1 Kinase Enzyme System can be purchased with or without the ADP-Glo™ Kinase Assay reagents. Used together, the ADP-Glo™ Kinase Assay + Kinase Enzyme Systems provide a convenient method for profiling the effect of lead compounds on kinase activity. Assay advantages include broad dynamic range, ease of use and high sensitivity. Kinase Enzyme Systems are manufactured by SignalChem. Bulk quantities available upon request.
Use with ADP-Glo™ Kinase Assay
The ADP-Glo™ Kinase Assay is a luminescent kinase assay that measures ADP formed from a kinase reaction; ADP is converted into ATP, which is a substrate in a reaction catalyzed by Ultra-Glo™ Luciferase that produces light. The luminescent signal positively correlates with ADP amount and kinase activity. The assay is well suited for measuring the effects of chemical compounds on the activity of a broad range of purified kinases, making it ideal for both primary screening as well as kinase selectivity profiling. The ADP-Glo™ Kinase Assay can be used to monitor the activity of virtually any ADP-generating enzyme (e.g., kinase or ATPase) using up to 1mM ATP.