PMID- 10653583
OWN - NLM
STAT- MEDLINE
DCOM- 20000210
LR  - 20181130
IS  - 0022-1554 (Print)
IS  - 0022-1554 (Linking)
VI  - 48
IP  - 1
DP  - 2000 Jan
TI  - Immunocytochemical localization of Shc and activated EGF receptor in early
      endosomes after EGF stimulation of HeLa cells.
PG  - 21-33
AB  - After binding of epidermal growth factor (EGF), the EGF receptor (EGFR) becomes
      autophosphorylated via tyrosine. The ligand-activated receptor is internalized by
      endocytosis and subsequently degraded in the lysosomal pathway. To follow EGFR
      activation after EGF stimulation, we generated antisera to the EGFR
      phosphotyrosine sites pY992 and pY1173. The SH2 region of Shc binds to both these
      sites. Both antisera identified EGFR after EGF binding and did not crossreact
      with the unactivated receptor. The intracellular distribution of phosphorylated
      EGFR after ligand binding was traced by two-color immunofluorescence confocal
      microscopy and immunoelectron microscopy. Before EGF stimulation EGFR was
      primarily located along the cell surface. When internalization of activated EGFR 
      was inhibited by incubation with EGF on ice, Y992- and Y1173-phosphorylated EGFR 
      were located along the plasma membrane. Ten minutes after internalization at 37C,
      Y992- and Y1173-phosphorylated EGFR were almost exclusively located in early
      endosomes, as shown by co-localization with EEA1. Immunoelectron microscopy
      confirmed that phosphorylated EGFR was located in intracellular vesicles
      resembling early endosomes. After EGF stimulation, the adaptor protein Shc
      redistributed to EGFR-containing early endosomes. Our results indicate that EGFR 
      activation of Shc via tyrosine-phosphorylated Y992 and Y1173 occurred in early
      endocytic compartments, and support a role for membrane trafficking in
      intracellular signaling.
FAU - Oksvold, M P
AU  - Oksvold MP
AD  - Laboratory for Toxicopathology, Institute of Pathology, The National Hospital,
      University of Oslo, Norway.
FAU - Skarpen, E
AU  - Skarpen E
FAU - Lindeman, B
AU  - Lindeman B
FAU - Roos, N
AU  - Roos N
FAU - Huitfeldt, H S
AU  - Huitfeldt HS
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Histochem Cytochem
JT  - The journal of histochemistry and cytochemistry : official journal of the
      Histochemistry Society
JID - 9815334
RN  - 0 (Adaptor Proteins, Signal Transducing)
RN  - 0 (Adaptor Proteins, Vesicular Transport)
RN  - 0 (Phosphoproteins)
RN  - 0 (Proteins)
RN  - 0 (SHC1 protein, human)
RN  - 0 (Shc Signaling Adaptor Proteins)
RN  - 0 (Src Homology 2 Domain-Containing, Transforming Protein 1)
RN  - 62229-50-9 (Epidermal Growth Factor)
RN  - EC 2.7.10.1 (ErbB Receptors)
SB  - IM
EIN - J Histochem Cytochem. 2017 Jul;65(7):421. PMID: 28651473
MH  - *Adaptor Proteins, Signal Transducing
MH  - *Adaptor Proteins, Vesicular Transport
MH  - Binding, Competitive
MH  - Biological Transport
MH  - Cell Compartmentation
MH  - Cell Membrane/metabolism
MH  - Endosomes/*ultrastructure
MH  - Enzyme-Linked Immunosorbent Assay
MH  - Epidermal Growth Factor/*pharmacology
MH  - ErbB Receptors/*isolation & purification
MH  - Fluorescent Antibody Technique
MH  - HeLa Cells
MH  - Humans
MH  - Intracellular Membranes/metabolism
MH  - Microscopy, Immunoelectron
MH  - Phosphoproteins/*isolation & purification
MH  - Phosphorylation
MH  - Proteins/*isolation & purification
MH  - Shc Signaling Adaptor Proteins
MH  - Src Homology 2 Domain-Containing, Transforming Protein 1
EDAT- 2000/02/01 00:00
MHDA- 2000/02/01 00:01
CRDT- 2000/02/01 00:00
PHST- 2000/02/01 00:00 [pubmed]
PHST- 2000/02/01 00:01 [medline]
PHST- 2000/02/01 00:00 [entrez]
AID - 10.1177/002215540004800103 [doi]
PST - ppublish
SO  - J Histochem Cytochem. 2000 Jan;48(1):21-33. doi: 10.1177/002215540004800103.