Cell Signaling Technology

Product Pathways - Apoptosis / Autophagy

DAPK1 Kinase #7647

Cell Signaling Technology offers a full line of protein kinases, substrates, antibody detection reagents and HTScan® kits. Browse our "Reagents for High-Throughput Screening" product listing or contact us at drugdiscovery@cellsignal.com.

Description

Purified recombinant kinase, supplied as a GST fusion protein.

Source / Purification

This GST-kinase fusion protein was produced using a baculovirus expression system with a construct expressing the kinase with an amino terminal GST tag. The protein was purified by one-step affinity chromatography using glutathione-agarose.

Quality Control

The purified kinase fusion protein was quality controlled for purity using SDS-PAGE followed by Coomassie or silver stain and Western blot. The specific activity of the kinase was determined using a radiometric assay.

Background

Death-associated protein kinase (DAPK) is a Ca2+/calmodulin-regulated serine/threonine kinase that participates in a wide range of apoptotic signals including interferon-γ, tumor necrosis factor α, Fas, activated c-Myc, and detachment from the extracellular matrix. In addition to the kinase domain and calmodulin regulatory segment, DAPK also has eight ankyrin repeats, a cytoskeleton binding region, and a conserved death domain (1-3). Deletion of the calmodulin-regulatory domain generates a constitutively active mutant kinase. Ectopic expression of wild-type DAPK induced cell death in HeLa cells. Conversely, expression of a catalytically inactive mutant protected cells from interferon-γ-induced cell death (4). The catalytic domain of DAPK has very high sequence similarity to vertebrate myosin light chain kinase (MLCK) and a RXX(S/T)X motif derived from myosin light chain protein was shown to be phosphorylated in vitro by DAPK (5).

  1. Kimchi, A. (1999) Ann Rheum Dis. 58, I14-I19.
  2. Cohen, O. et al. (1999) J Cell Biol 146, 141-148.
  3. Deiss, L. P. et al. (1995) Genes Dev 9, 15-30.
  4. Cohen, O. et al. (1997) EMBO J 16, 998-1008.
  5. Velentza, A. V. et al. (2001) J Biol Chem 276, 38956-38965.

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