PMID- 8700533
OWN - NLM
STAT- MEDLINE
DCOM- 19960904
LR  - 20211203
IS  - 0950-9232 (Print)
IS  - 0950-9232 (Linking)
VI  - 12
IP  - 12
DP  - 1996 Jun 20
TI  - HER-2/c-erbB2 is phosphorylated by calmodulin-dependent protein kinase II on a 
      single site in the cytoplasmic tail at threonine-1172.
PG  - 2725-30
AB  - Calmodulin-dependent protein kinase II (Cam kinase II) is known to desensitise 
      epidermal growth factor receptor (HER-1) tyrosine kinase activity by a process 
      involving phosphorylation at serines 1046/47 in the cytoplasmic tail. We have 
      developed an experimental system to investigate phosphorylation of the related 
      HER-2/c-erbB2 proto-oncogene utilising purified Cam kinase II and recombinant 
      glutathione-S-transferase (GST) fusion proteins. The cDNA for rat Cam kinase 
      II-alpha was transfected into human embryonic kidney (HEK) 293 fibroblasts and 
      the expressed protein purified to homogeneity by calmodulin-agarose affinity 
      chromatography. A GST fusion protein comprising residues 1126-1255 of HER-2 was 
      phosphorylated by purified Cam kinase II, in contrast to a GST protein comprising 
      residues 1005-1125. Phosphoamino-acid analysis and site-directed mutagenesis 
      indicated that HER-2 was phosphorylated on a single site at threonine-1172 which 
      resides within a consensus Cam kinase II phosphorylation site (RAKT). HER-2 
      (threonine-1172-alanine), in the form of a ligand-inducible chimaera HER-1/2, was 
      co-transfected into HEK-293 fibroblasts with a constitutively active form of Cam 
      kinase II, followed by in vivo labelling of these cells with 32 P-orthophosphate. 
      Immunoprecipitation of ligand-activated receptors followed by two-dimensional 
      phosphopeptide mapping indicated that threonine-1172 in HER-2 is a newly 
      identified in vivo site which can be hyper-phosphorylated by constitutively 
      active Cam kinase II. In addition, when over-expressed in HEK-293 fibroblasts, 
      HER-1/2 (threonine-1172-alanine) showed a defect in desensitisation and underwent 
      a more sustained EGF-induced receptor autophosphorylation compared to wild-type 
      HER-1/2.
FAU - Feinmesser, R L
AU  - Feinmesser RL
AD  - Department of Biochemistry, Charing Cross and Westminster Medical School, London, 
      UK.
FAU - Gray, K
AU  - Gray K
FAU - Means, A R
AU  - Means AR
FAU - Chantry, A
AU  - Chantry A
LA  - eng
GR  - Wellcome Trust/United Kingdom
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Oncogene
JT  - Oncogene
JID - 8711562
RN  - 0 (MAS1 protein, human)
RN  - 0 (Phosphopeptides)
RN  - 0 (Proto-Oncogene Mas)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 2ZD004190S (Threonine)
RN  - 62229-50-9 (Epidermal Growth Factor)
RN  - EC 2.7.10.1 (ErbB Receptors)
RN  - EC 2.7.10.1 (Receptor, ErbB-2)
RN  - EC 2.7.11.17 (CAMK2A protein, human)
RN  - EC 2.7.11.17 (Calcium-Calmodulin-Dependent Protein Kinase Type 2)
RN  - EC 2.7.11.17 (Calcium-Calmodulin-Dependent Protein Kinases)
RN  - EC 2.7.11.17 (Camk2a protein, rat)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Binding Sites
MH  - Calcium-Calmodulin-Dependent Protein Kinase Type 2
MH  - Calcium-Calmodulin-Dependent Protein Kinases/*metabolism
MH  - Cytoplasm/*chemistry
MH  - Epidermal Growth Factor/pharmacology
MH  - ErbB Receptors/chemistry/metabolism
MH  - Fibroblasts/drug effects
MH  - Humans
MH  - Molecular Sequence Data
MH  - Phosphopeptides/biosynthesis/genetics
MH  - Phosphorylation
MH  - Proto-Oncogene Mas
MH  - Rats
MH  - Receptor, ErbB-2/*metabolism
MH  - Recombinant Fusion Proteins/genetics/metabolism
MH  - Sequence Homology, Amino Acid
MH  - Substrate Specificity
MH  - Threonine/*metabolism
EDAT- 1996/06/20 00:00
MHDA- 2000/03/23 09:00
CRDT- 1996/06/20 00:00
PHST- 1996/06/20 00:00 [pubmed]
PHST- 2000/03/23 09:00 [medline]
PHST- 1996/06/20 00:00 [entrez]
PST - ppublish
SO  - Oncogene. 1996 Jun 20;12(12):2725-30.