PMID- 10336483
OWN - NLM
STAT- MEDLINE
DCOM- 19990629
LR  - 20211203
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 274
IP  - 22
DP  - 1999 May 28
TI  - Substrate recognition by Ca2+/Calmodulin-dependent protein kinase kinase. Role of
      the arg-pro-rich insert domain.
PG  - 15803-10
AB  - Mammalian Ca2+/CaM-dependent protein kinase kinase (CaM-KK) has been identified
      and cloned as an activator for two kinases, CaM kinase I (CaM-KI) and CaM kinase 
      IV (CaM-KIV), and a recent report (Yano, S., Tokumitsu, H., and Soderling, T. R. 
      (1998) Nature 396, 584-587) demonstrates that CaM-KK can also activate and
      phosphorylate protein kinase B (PKB). In this study, we identify a CaM-KK from
      Caenorhabditis elegans, and comparison of its sequence with the mammalian CaM-KK 
      alpha and beta shows a unique Arg-Pro (RP)-rich insert in their catalytic domains
      relative to other protein kinases. Deletion of the RP-domain resulted in complete
      loss of CaM-KIV activation activity and physical interaction of CaM-KK with
      glutathione S-transferase-CaM-KIV (T196A). However, CaM-KK autophosphorylation
      and phosphorylation of a synthetic peptide substrate were normal in the RP-domain
      mutant. Site-directed mutagenesis of three conserved Arg in the RP- domain of
      CaM-KK confirmed that these positive charges are important for CaM-KIV
      activation. The RP- domain deletion mutant also failed to fully activate and
      phosphorylate CaM-KI, but this mutant was indistinguishable from wild-type CaM-KK
      for the phosphorylation and activation of PKB. These results indicate that the
      RP-domain in CaM-KK is critical for recognition of downstream CaM-kinases but not
      for its catalytic activity (i.e. autophosphorylation) and PKB activation.
FAU - Tokumitsu, H
AU  - Tokumitsu H
AD  - Helix Research Institute, Inc., 1532-3 Yana, Kisarazu-shi, Chiba 292-0812, Japan.
FAU - Takahashi, N
AU  - Takahashi N
FAU - Eto, K
AU  - Eto K
FAU - Yano, S
AU  - Yano S
FAU - Soderling, T R
AU  - Soderling TR
FAU - Muramatsu, M
AU  - Muramatsu M
LA  - eng
SI  - GENBANK/AB016838
PT  - Journal Article
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Helminth Proteins)
RN  - 0 (Proto-Oncogene Proteins)
RN  - 0 (Recombinant Proteins)
RN  - EC 2.7.11.1 (Protein Serine-Threonine Kinases)
RN  - EC 2.7.11.1 (Proto-Oncogene Proteins c-akt)
RN  - EC 2.7.11.17 (Calcium-Calmodulin-Dependent Protein Kinase Kinase)
RN  - EC 2.7.11.17 (Calcium-Calmodulin-Dependent Protein Kinase Type 4)
RN  - EC 2.7.11.17 (Calcium-Calmodulin-Dependent Protein Kinases)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Base Sequence
MH  - Binding Sites
MH  - Caenorhabditis elegans/*enzymology
MH  - Calcium-Calmodulin-Dependent Protein Kinase Kinase
MH  - Calcium-Calmodulin-Dependent Protein Kinase Type 4
MH  - Calcium-Calmodulin-Dependent Protein Kinases/chemistry/*genetics/metabolism
MH  - Cloning, Molecular
MH  - Enzyme Activation
MH  - Helminth Proteins/chemistry/genetics
MH  - Molecular Sequence Data
MH  - Mutagenesis, Site-Directed
MH  - Phosphorylation
MH  - Protein Serine-Threonine Kinases/chemistry
MH  - Proto-Oncogene Proteins/metabolism
MH  - Proto-Oncogene Proteins c-akt
MH  - Recombinant Proteins/metabolism
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - Sequence Deletion
MH  - Sequence Homology, Amino Acid
EDAT- 1999/05/21 00:00
MHDA- 1999/05/21 00:01
CRDT- 1999/05/21 00:00
PHST- 1999/05/21 00:00 [pubmed]
PHST- 1999/05/21 00:01 [medline]
PHST- 1999/05/21 00:00 [entrez]
AID - 10.1074/jbc.274.22.15803 [doi]
AID - S0021-9258(19)73092-0 [pii]
PST - ppublish
SO  - J Biol Chem. 1999 May 28;274(22):15803-10. doi: 10.1074/jbc.274.22.15803.