PMID- 15500439
OWN - NLM
STAT- MEDLINE
DCOM- 20050715
LR  - 20181113
IS  - 1470-8728 (Electronic)
IS  - 0264-6021 (Print)
IS  - 0264-6021 (Linking)
VI  - 387
IP  - Pt 1
DP  - 2005 Apr 1
TI  - Distinct signalling particles containing ERK/MEK and B-Raf in PC12 cells.
PG  - 155-64
AB  - Although several multiprotein complexes containing MAPKs (mitogen-activated 
      protein kinases) have been identified using overexpression of kinases and 
      scaffold proteins, the components of the complexes and their physical properties 
      at endogenous expression levels have not been defined. We characterized a large 
      protein complex containing a nerve-growth-factor-activated ERK 
      (extracellular-signal-regulated kinase) and MEK (MAPK/ERK kinase) in rat 
      pheochromocytoma (PC12) cells. This protein complex fractionated into a 
      high-speed pellet and was resistant to non-ionic detergent treatments that 
      solubilized membranes. Disruption of protein-protein interactions by treatment 
      with high salt was required to facilitate immunoprecipitation of active ERK1 and 
      co-precipitation of MEK1. Microtubule fragments were also present in the 
      detergent-resistant high-speed pellet, and some kinases were bound to them, 
      especially ERK1b (an alternatively spliced isoform of ERK1), which showed a 
      strong preference for binding microtubules. The large protein complex containing 
      ERK1 and MEK1 was resolved by velocity sedimentation from fragments of 
      microtubules; however, it did not contain other scaffolding components known to 
      bind ERK and MEK. B-Raf was also present in a distinct detergent-resistant, 
      microtubule-independent protein complex slightly larger than that containing ERK 
      and MEK. We conclude that there are two independent nerve growth factor-regulated 
      'signalling particles' with an estimated size of 60-75 S, one containing ERK1 and 
      MEK1 and the other containing B-Raf. These signalling particles may have a role 
      in the temporal and spatial regulation of kinase activity inside cells.
FAU - MacCormick, Matt
AU  - MacCormick M
AD  - Institute of Molecular Biosciences, Massey University, Private Bag 11222, 
      Palmerston North, New Zealand.
FAU - Moderscheim, Tanja
AU  - Moderscheim T
FAU - van der Salm, Louise W M
AU  - van der Salm LW
FAU - Moore, Anna
AU  - Moore A
FAU - Pryor, Shona Clements
AU  - Pryor SC
FAU - McCaffrey, Gretchen
AU  - McCaffrey G
FAU - Grimes, Mark L
AU  - Grimes ML
LA  - eng
GR  - P20 RR015583/RR/NCRR NIH HHS/United States
GR  - P20 RR15583/RR/NCRR NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - England
TA  - Biochem J
JT  - The Biochemical journal
JID - 2984726R
RN  - 0 (Isoenzymes)
RN  - EC 2.7.11.1 (Proto-Oncogene Proteins B-raf)
RN  - EC 2.7.11.24 (Extracellular Signal-Regulated MAP Kinases)
RN  - EC 2.7.11.24 (Mitogen-Activated Protein Kinase 3)
RN  - EC 2.7.11.24 (Mitogen-Activated Protein Kinases)
SB  - IM
MH  - Alternative Splicing/physiology
MH  - Animals
MH  - Cell Fractionation/methods
MH  - Extracellular Signal-Regulated MAP Kinases/chemistry/*metabolism
MH  - Isoenzymes/metabolism
MH  - MAP Kinase Signaling System/physiology
MH  - Microtubules/chemistry/metabolism
MH  - Mitogen-Activated Protein Kinase 3/metabolism
MH  - Mitogen-Activated Protein Kinases/*metabolism
MH  - PC12 Cells/*chemistry/*metabolism
MH  - Proto-Oncogene Proteins B-raf/*metabolism
MH  - Rats
MH  - Signal Transduction/*physiology
PMC - PMC1134943
EDAT- 2004/10/27 09:00
MHDA- 2005/07/16 09:00
PMCR- 2005/10/01
CRDT- 2004/10/27 09:00
PHST- 2004/10/27 09:00 [pubmed]
PHST- 2005/07/16 09:00 [medline]
PHST- 2004/10/27 09:00 [entrez]
PHST- 2005/10/01 00:00 [pmc-release]
AID - BJ20040272 [pii]
AID - bj3870155 [pii]
AID - 10.1042/BJ20040272 [doi]
PST - ppublish
SO  - Biochem J. 2005 Apr 1;387(Pt 1):155-64. doi: 10.1042/BJ20040272.