PMID- 12861023
OWN - NLM
STAT- MEDLINE
DCOM- 20030829
LR  - 20211203
IS  - 0270-7306 (Print)
IS  - 1098-5549 (Electronic)
IS  - 0270-7306 (Linking)
VI  - 23
IP  - 15
DP  - 2003 Aug
TI  - Proteomic identification of 14-3-3zeta as a mitogen-activated protein 
      kinase-activated protein kinase 2 substrate: role in dimer formation and ligand 
      binding.
PG  - 5376-87
AB  - Mitogen-activated protein kinase (MAPK)-activated protein kinase 2 (MAPKAPK2) 
      mediates multiple p38 MAPK-dependent inflammatory responses. To define the signal 
      transduction pathways activated by MAPKAPK2, we identified potential MAPKAPK2 
      substrates by using a functional proteomic approach consisting of in vitro 
      phosphorylation of neutrophil lysate by active recombinant MAPKAPK2, protein 
      separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis 
      (SDS-PAGE), and phosphoprotein identification by peptide mass fingerprinting with 
      matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS) and 
      protein database analysis. One of the eight candidate MAPKAPK2 substrates 
      identified was the adaptor protein, 14-3-3zeta. We confirmed that MAPKAPK2 
      interacted with and phosphorylated 14-3-3zeta in vitro and in HEK293 cells. The 
      chemoattractant formyl-methionyl-leucyl-phenylalanine (fMLP) stimulated 
      p38-MAPK-dependent phosphorylation of 14-3-3 proteins in human neutrophils. 
      Mutation analysis showed that MAPKAPK2 phosphorylated 14-3-3zeta at Ser-58. 
      Computational modeling and calculation of theoretical binding energies predicted 
      that both phosphorylation at Ser-58 and mutation of Ser-58 to Asp (S58D) 
      compromised the ability of 14-3-3zeta to dimerize. Experimentally, S58D mutation 
      significantly impaired both 14-3-3zeta dimerization and binding to Raf-1. These 
      data suggest that MAPKAPK2-mediated phosphorylation regulates 14-3-3zeta 
      functions, and this MAPKAPK2 activity may represent a novel pathway mediating p38 
      MAPK-dependent inflammation.
FAU - Powell, David W
AU  - Powell DW
AD  - Departments of Biochemistry and Molecular Biology, University of Louisville, 
      Louisville, KY 40202, USA.
FAU - Rane, Madhavi J
AU  - Rane MJ
FAU - Joughin, Brian A
AU  - Joughin BA
FAU - Kalmukova, Ralitsa
AU  - Kalmukova R
FAU - Hong, Jeong-Ho
AU  - Hong JH
FAU - Tidor, Bruce
AU  - Tidor B
FAU - Dean, William L
AU  - Dean WL
FAU - Pierce, William M
AU  - Pierce WM
FAU - Klein, Jon B
AU  - Klein JB
FAU - Yaffe, Michael B
AU  - Yaffe MB
FAU - McLeish, Kenneth R
AU  - McLeish KR
LA  - eng
GR  - R01 HL 66358/HL/NHLBI NIH HHS/United States
GR  - R01 GM059281/GM/NIGMS NIH HHS/United States
GR  - R01 GM060594/GM/NIGMS NIH HHS/United States
GR  - R01 HL066358/HL/NHLBI NIH HHS/United States
GR  - R01 GM 60594/GM/NIGMS NIH HHS/United States
GR  - R21 DK 06289/DK/NIDDK NIH HHS/United States
GR  - R01 GM 59281/GM/NIGMS NIH HHS/United States
GR  - R21 DK 62086/DK/NIDDK NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Mol Cell Biol
JT  - Molecular and cellular biology
JID - 8109087
RN  - 0 (14-3-3 Proteins)
RN  - 0 (Intracellular Signaling Peptides and Proteins)
RN  - 0 (Ligands)
RN  - 0 (Proteome)
RN  - 0 (Recombinant Proteins)
RN  - 59880-97-6 (N-Formylmethionine Leucyl-Phenylalanine)
RN  - EC 1.14.16.2 (Tyrosine 3-Monooxygenase)
RN  - EC 2.5.1.18 (Glutathione Transferase)
RN  - EC 2.7.1.- (MAP-kinase-activated kinase 2)
RN  - EC 2.7.11.1 (Protein Serine-Threonine Kinases)
RN  - EC 2.7.11.1 (Proto-Oncogene Proteins c-raf)
RN  - EC 2.7.11.24 (Mitogen-Activated Protein Kinases)
RN  - EC 2.7.11.24 (p38 Mitogen-Activated Protein Kinases)
RN  - EC 3.4.21.4 (Trypsin)
SB  - IM
MH  - 14-3-3 Proteins
MH  - Amino Acid Sequence
MH  - Cell Line
MH  - DNA Mutational Analysis
MH  - Dimerization
MH  - Electrophoresis, Polyacrylamide Gel
MH  - Genetic Vectors
MH  - Glutathione Transferase/metabolism
MH  - Humans
MH  - Intracellular Signaling Peptides and Proteins
MH  - Ligands
MH  - *MAP Kinase Signaling System
MH  - Mitogen-Activated Protein Kinases/metabolism
MH  - Models, Molecular
MH  - Molecular Sequence Data
MH  - Mutation
MH  - N-Formylmethionine Leucyl-Phenylalanine/pharmacology
MH  - Neutrophils/enzymology/metabolism
MH  - Phosphorylation
MH  - Precipitin Tests
MH  - Protein Binding
MH  - Protein Serine-Threonine Kinases/*metabolism
MH  - Proteome
MH  - Proto-Oncogene Proteins c-raf/metabolism
MH  - Recombinant Proteins/metabolism
MH  - Signal Transduction
MH  - Software
MH  - Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
MH  - Substrate Specificity
MH  - Trypsin/pharmacology
MH  - Tyrosine 3-Monooxygenase/chemistry/*physiology
MH  - p38 Mitogen-Activated Protein Kinases
PMC - PMC165733
EDAT- 2003/07/16 05:00
MHDA- 2003/08/30 05:00
PMCR- 2003/08/01
CRDT- 2003/07/16 05:00
PHST- 2003/07/16 05:00 [pubmed]
PHST- 2003/08/30 05:00 [medline]
PHST- 2003/07/16 05:00 [entrez]
PHST- 2003/08/01 00:00 [pmc-release]
AID - 1707 [pii]
AID - 10.1128/MCB.23.15.5376-5387.2003 [doi]
PST - ppublish
SO  - Mol Cell Biol. 2003 Aug;23(15):5376-87. doi: 10.1128/MCB.23.15.5376-5387.2003.