kinase D (PKD), a serine/threonine kinase originally described as a novel
PKC family member and termed PKCm
(1), belongs to the calcium calmodulin superfamily of kinases (2-3). Three
mammalian isoforms have so far been described - PKD1/PKCm, PKD2 and PKD3/PKCn; these isoforms show a
high degree of homology, especially in their catalytic domain
(4-5). PKDs are major targets for tumor promoting
esters; they are activated via G protein-coupled receptors (GPCRs) and their activation is also dependent on PKC activation
(5). PKDs have been implicated
in numerous cellular functions, including signal transduction as well as cell
survival, migration, differentiation, and proliferation (4-6). They are important
regulators of secretory transport at the trans-golgi network (7). Of the three isoforms, PKD1 is the best characterized.
It is involved in the regulation of Golgi function, cell proliferation and
apoptosis (8) and it mediates oxidative stress signaling regulating cellular
detoxification and survival (9). PKD2 has been found to phosphorylate histone
H1 more efficiently than aldolase in vitro (10).
is pleased to announce the release of isoform specific PKD1 and PKD2 antibodies.
These antibodies were raised using sequences from the C-terminus of PKD1 and
PKD2. Anti-PKD1 only recognizes PKD1 and not PKD2 and PKD3. Anti-PKD2 only
recognizes PKD2 and not PKD1 or PKD3.
||Figure 1: Western blots: HEK293T cells
were transfected with HA-tagged human PKD1, Flag-tagged human PKD2 and
GST-tagged human PKD3 and incubated at 37°C for 24 hours. The cultures
were then lysed and run on a 7.5% SDS-PAGE gel. Blots probed with anti-HA,
-Flag (anti-DYKDDDDK Tag) and anti-GST at a final concentration of 200
ng/ml in 5% milk in TBST at 4°C overnight. Courtesy of Dr. Alex Toker,
Harvard Medical School, Boston, MA.
||Figure 2. Western
blots: HEK293T cells were transfected with HA-tagged human PKD1, Flag-tagged
human PKD2 and GST-tagged human PKD3 and incubated at 37°C for 24 hours.
The cultures were then lysed and 100 ug of total lysate run on a 7.5%
SDS-PAGE gel. Blots were then probed with -human PKD2 kinase (Cat# 55142),
and control -HA, anti-Flag (anti-DYKDDDDK Tag) and -GST at a final concentration
of 200 ng/ml in 5% milk in TBST at 4°C overnight. Courtesy of Dr. Alex
Toker, Harvard Medical School, Boston, MA.
Protein Kinase antibodies.
of kinase/phosphatase peptide substrates
1. Johannes, FJ. et al. J.
Biol. Chem. 269, 6140 (1994).
2. Manning, G. et al. Science
298, 1912 (2002).
3. Van Lint, J. et al. Trends Cell
Biol. 12, 193 (2002).
4. Waldron, RT. et al. J. Biol.
Chem. 276, 32606 (2001).
5. Auer, A. et al. Mol. Bio.
Cell. 16, 4375 (2005).
6. Rozengurt, E. et al. J. Biol.
Chem. 208, 13205 (2005).
7. Yeaman, C. et al. Nature
Cell Biol. 6, 106 (2004).
8. Rykx, A. et al. FEBS Lett.
546, 81 (2003).
H. & P. Storz, et al. J. Biol. Chem. 282,
10.Sturany, S. et al. J.
Biol. Chem. 276, 3310 (2001).