PMID- 10197446
OWN - NLM
STAT- MEDLINE
DCOM- 19990622
LR  - 20071115
IS  - 0173-0835 (Print)
IS  - 0173-0835 (Linking)
VI  - 20
IP  - 2
DP  - 1999 Feb
TI  - Phosphorylation-dependent protein kinase D activation.
PG  - 382-90
AB  - The novel mouse serine-threonine kinase protein kinase D (PKD) is activated in
      intact Swiss 3T3 cells stimulated by phorbol esters, cell permeant
      diacylglycerols, bryostatin, neuropeptides and growth factors via a
      phosphorylation-dependent mechanism requiring protein kinase C (PKC) activity.
      Structural comparison of the PKD catalytic domain with other kinases reveals a
      close similarity with MEK family kinases, which are activated upon
      phosphorylation of key serine and threonine residues in a region termed the
      activation loop. To study the regulation of PKD, we transfected mutant PKD cDNAs 
      in which putative activation loop serine residues 744 and 748 were mutated to
      either alanine or glutamic acid into COS-7 cells. Replacement of serines 744 and 
      748 with alanine prevented activation of the overexpressed PKD form upon phorbol 
      ester treatment of cells, whereas replacement with glutamic acid results in full 
      constitutive activation. Single serine to glutamic acid replacement mutants were 
      partially activated. In vivo 32P-labeling and two-dimensional phosphopeptide
      mapping of PKD and catalytically inactive PKD mutants at serine 744, 748 or at
      both residues revealed that phorbol ester-sensitive phosphopeptides could be
      selectively eliminated from patterns observed as a result of these mutations.
      Treatment of cells with the PKC inhibitor GFI also prevented the appearance of
      phosphopeptide spots occuring in response to phorbol ester stimulation. These
      results provide direct evidence that PKD becomes activated in vivo as a
      consequence of PKC-mediated phosphorylation of serines 744 and 748. These results
      support our view of PKD as an important clement in PKC signal transduction.
FAU - Waldron, R T
AU  - Waldron RT
AD  - Department of Medicine, School of Medicine and Molecular Biology, University of
      California, Los Angeles 90095-1786, USA.
FAU - Iglesias, T
AU  - Iglesias T
FAU - Rozengurt, E
AU  - Rozengurt E
LA  - eng
PT  - Journal Article
PT  - Review
PL  - Germany
TA  - Electrophoresis
JT  - Electrophoresis
JID - 8204476
RN  - EC 2.7.10.- (protein kinase D)
RN  - EC 2.7.11.13 (Protein Kinase C)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Binding Sites
MH  - Enzyme Activation
MH  - Humans
MH  - Molecular Sequence Data
MH  - Phosphorylation
MH  - Protein Kinase C/*metabolism
RF  - 54
EDAT- 1999/04/10 02:04
MHDA- 2000/08/12 11:00
CRDT- 1999/04/10 02:04
PHST- 1999/04/10 02:04 [pubmed]
PHST- 2000/08/12 11:00 [medline]
PHST- 1999/04/10 02:04 [entrez]
AID - 10.1002/(SICI)1522-2683(19990201)20:2<382::AID-ELPS382>3.0.CO;2-N [pii]
AID - 10.1002/(SICI)1522-2683(19990201)20:2<382::AID-ELPS382>3.0.CO;2-N [doi]
PST - ppublish
SO  - Electrophoresis. 1999 Feb;20(2):382-90. doi:
      10.1002/(SICI)1522-2683(19990201)20:2<382::AID-ELPS382>3.0.CO;2-N.