PMID- 10618391
OWN - NLM
STAT- MEDLINE
DCOM- 20000210
LR  - 20190501
IS  - 0027-8424 (Print)
IS  - 0027-8424 (Linking)
VI  - 97
IP  - 1
DP  - 2000 Jan 4
TI  - Analysis of receptor signaling pathways by mass spectrometry: identification of
      vav-2 as a substrate of the epidermal and platelet-derived growth factor
      receptors.
PG  - 179-84
AB  - Oligomerization of receptor protein tyrosine kinases such as the epidermal growth
      factor receptor (EGFR) by their cognate ligands leads to activation of the
      receptor. Transphosphorylation of the receptor subunits is followed by the
      recruitment of signaling molecules containing src homology 2 (SH2) or
      phosphotyrosine interaction domains (PID). Additionally, several cytoplasmic
      proteins that may or may not associate with the receptor undergo tyrosine
      phosphorylation. To identify several components of the EGFR signaling pathway in 
      a single step, we have immunoprecipitated molecules that are tyrosine
      phosphorylated in response to EGF and analyzed them by one-dimensional gel
      electrophoresis followed by mass spectrometry. Combining matrix-assisted laser
      desorption/ionization (MALDI) and nanoelectrospray tandem mass spectrometry
      (MS/MS) led to the identification of nine signaling molecules, seven of which had
      previously been implicated in EGFR signaling. Several of these molecules were
      identified from low femtomole levels of protein loaded onto the gel. We
      identified Vav-2, a recently discovered guanosine nucleotide exchange factor that
      is expressed ubiquitously, as a substrate of the EGFR. We demonstrate that Vav-2 
      is phosphorylated on tyrosine residues in response to EGF and associates with the
      EGFR in vivo. Binding of Vav-2 to the EGFR is mediated by the SH2 domain of
      Vav-2. In keeping with its ubiquitous expression, Vav-2 seems to be a general
      signaling molecule, since it also associates with the platelet-derived growth
      factor (PDGF) receptor and undergoes tyrosine phosphorylation in fibroblasts upon
      PDGF stimulation. The strategy suggested here can be used for routine
      identification of downstream components of cell surface receptors in mammalian
      cells.
FAU - Pandey, A
AU  - Pandey A
AD  - Whitehead Institute for Biomedical Research, Cambridge, MA 02142, USA.
FAU - Podtelejnikov, A V
AU  - Podtelejnikov AV
FAU - Blagoev, B
AU  - Blagoev B
FAU - Bustelo, X R
AU  - Bustelo XR
FAU - Mann, M
AU  - Mann M
FAU - Lodish, H F
AU  - Lodish HF
LA  - eng
GR  - P01 HL032262/HL/NHLBI NIH HHS/United States
GR  - K01 CA75447/CA/NCI NIH HHS/United States
GR  - P01 HL32262/HL/NHLBI 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  - Proc Natl Acad Sci U S A
JT  - Proceedings of the National Academy of Sciences of the United States of America
JID - 7505876
RN  - 0 (Oncogene Proteins)
RN  - 0 (Proto-Oncogene Proteins c-vav)
RN  - 0 (VAV2 protein, human)
RN  - 21820-51-9 (Phosphotyrosine)
RN  - EC 2.7.10.1 (ErbB Receptors)
RN  - EC 2.7.10.1 (Receptor Protein-Tyrosine Kinases)
RN  - EC 2.7.10.1 (Receptors, Platelet-Derived Growth Factor)
SB  - IM
MH  - ErbB Receptors/*metabolism
MH  - HeLa Cells
MH  - Humans
MH  - Mass Spectrometry/methods
MH  - Oncogene Proteins/*metabolism
MH  - Phosphorylation
MH  - Phosphotyrosine/metabolism
MH  - Precipitin Tests
MH  - Protein Binding
MH  - Proto-Oncogene Proteins c-vav
MH  - Receptor Protein-Tyrosine Kinases/metabolism
MH  - Receptors, Platelet-Derived Growth Factor/*metabolism
MH  - *Signal Transduction
MH  - src Homology Domains
PMC - PMC26636
EDAT- 2000/01/05 00:00
MHDA- 2000/01/05 00:01
CRDT- 2000/01/05 00:00
PHST- 2000/01/05 00:00 [pubmed]
PHST- 2000/01/05 00:01 [medline]
PHST- 2000/01/05 00:00 [entrez]
AID - 10.1073/pnas.97.1.179 [doi]
PST - ppublish
SO  - Proc Natl Acad Sci U S A. 2000 Jan 4;97(1):179-84. doi: 10.1073/pnas.97.1.179.