PMID- 10551809
OWN - NLM
STAT- MEDLINE
DCOM- 20000103
LR  - 20210209
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 274
IP  - 46
DP  - 1999 Nov 12
TI  - Identification of a central phosphorylation site in p21-activated kinase
      regulating autoinhibition and kinase activity.
PG  - 32565-73
AB  - p21-activated kinases (Pak)/Ste20 kinases are regulated in vitro and in vivo by
      the small GTP-binding proteins Rac and Cdc42 and lipids, such as sphingosine,
      which stimulate autophosphorylation and phosphorylation of exogenous substrates. 
      The mechanism of Pak activation by these agents remains unclear. We investigated 
      Pak kinase activation in more detail to gain insight into the interplay between
      the GTPase/sphingosine binding, an intramolecular inhibitory interaction, and
      autophosphorylation. We present biochemical evidence that an autoinhibitory
      domain (ID) contained within amino acid residues 67-150 of Pak1 interacts with
      the carboxyl-terminal kinase domain and that this interaction is regulated in a
      GTPase-dependent fashion. Cdc42- and sphingosine-stimulated Pak1 activity can be 
      inhibited in trans by recombinant ID peptide, indicating similarities in their
      mode of activation. However, Pak1, which was autophosphorylated in response to
      either GTPase or sphingosine, is highly active and is insensitive to inhibition
      by the ID peptide. We identified phospho-acceptor site threonine 423 in the
      kinase activation loop as a critical determinant for the sensitivity to
      autoinhibition and enzymatic activity. Phosphorylation studies suggested that the
      stimulatory effect of both GTPase and sphingosine results in exposure of the
      activation loop, making it accessible for intermolecular phosphorylation.
FAU - Zenke, F T
AU  - Zenke FT
AD  - Department of Immunology, The Scripps Research Institute, La Jolla, California
      92037, USA.
FAU - King, C C
AU  - King CC
FAU - Bohl, B P
AU  - Bohl BP
FAU - Bokoch, G M
AU  - Bokoch GM
LA  - eng
GR  - AG15430/AG/NIA NIH HHS/United States
GR  - GM 39434/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Intracellular Signaling Peptides and Proteins)
RN  - 0 (Peptide Fragments)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Saccharomyces cerevisiae Proteins)
RN  - EC 2.7.11.1 (PAK1 protein, human)
RN  - EC 2.7.11.1 (Pak1 protein, rat)
RN  - EC 2.7.11.1 (Protein-Serine-Threonine Kinases)
RN  - EC 2.7.11.1 (p21-Activated Kinases)
RN  - EC 2.7.11.25 (MAP Kinase Kinase Kinases)
RN  - EC 2.7.11.25 (STE20 protein, S cerevisiae)
RN  - EC 3.6.1.- (GTP-Binding Proteins)
RN  - EC 3.6.5.2 (cdc42 GTP-Binding Protein)
RN  - NGZ37HRE42 (Sphingosine)
SB  - IM
MH  - Animals
MH  - Binding Sites
MH  - Enzyme Activation
MH  - GTP-Binding Proteins/metabolism
MH  - Humans
MH  - Intracellular Signaling Peptides and Proteins
MH  - MAP Kinase Kinase Kinases
MH  - Mutation
MH  - Peptide Fragments/pharmacology
MH  - Phosphorylation
MH  - Protein Conformation
MH  - Protein-Serine-Threonine Kinases/antagonists & inhibitors/genetics/*metabolism
MH  - Rats
MH  - Recombinant Proteins
MH  - *Saccharomyces cerevisiae Proteins
MH  - Sphingosine/pharmacology
MH  - cdc42 GTP-Binding Protein/metabolism
MH  - p21-Activated Kinases
EDAT- 1999/11/07 00:00
MHDA- 1999/11/07 00:01
CRDT- 1999/11/07 00:00
PHST- 1999/11/07 00:00 [pubmed]
PHST- 1999/11/07 00:01 [medline]
PHST- 1999/11/07 00:00 [entrez]
AID - 10.1074/jbc.274.46.32565 [doi]
AID - S0021-9258(17)46548-3 [pii]
PST - ppublish
SO  - J Biol Chem. 1999 Nov 12;274(46):32565-73. doi: 10.1074/jbc.274.46.32565.