The Kinase Enzyme Systems include a recombinant kinase enzyme, a substrate appropriate for the enzyme, a reaction buffer and supplemental reagents as needed. The MLCK Kinase Enzyme System contains:
- MLCK Kinase, 10μg (Human, recombinant; amino acids 1425–1776). MW: ~70kDa.
- MRCL3 Peptide (KKRPQRATSN-VFAM-NH2) Substrate; derived from human myosin regulatory light chain MRCL3 (amino acid 11–24).
- Reaction Buffer, DTT, Ca2+/Calmodulin Solution.
Recombinant human MLCK (1425–1776) was expressed by baculovirus in Sf9 insect cells using a N-terminal GST tag. MLCK or myosin light chain kinase is a muscle member of the immunoglobulin gene superfamily and is a calcium/calmodulin-dependent protein kinase. MLCK is a regulatory protein for smooth muscle contraction, which acts by phosphorylating 20-kDa myosin light chain (MLC20) to activate the myosin ATPase activity. The kinase activity for the phosphorylation is localized at the central part of MLCK, which is also furnished with actin-binding activity at its N terminal and myosin-binding activity at its C terminal. Phosphorylation of regulatory light chain of myosin plays an important role in controlling the morphological changes seen during cell division.
MLCK NCBI Database Entry.
The MLCK 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.
See all Kinase Enzyme Systems available from Promega.