PMID- 10497257
OWN - NLM
STAT- MEDLINE
DCOM- 19991102
LR  - 20190508
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 274
IP  - 40
DP  - 1999 Oct 1
TI  - The shedding of membrane-anchored heparin-binding epidermal-like growth factor is
      regulated by the Raf/mitogen-activated protein kinase cascade and by cell
      adhesion and spreading.
PG  - 28828-35
AB  - Heparin-binding epidermal-like growth factor (HB-EGF) is synthesized as a
      transmembrane precursor (HB-EGF(TM)). The addition of phorbol ester (PMA, phorbol
      12-myristate 13-acetate) to cells expressing HB-EGF(TM) results in the
      metalloproteinase-dependent release (shedding) of soluble HB-EGF. To analyze
      mechanisms that regulate HB-EGF shedding, a stable cell line was established
      expressing HB-EGF(TM) in which the ectodomain and the cytoplasmic tail were
      tagged with hemagglutinin (HA) and Myc epitopes, respectively (HB-EGF(TM)HA/Myc).
      HB-EGF(TM)HA/Myc cleavage was followed by the appearance of soluble HB-EGFHA in
      conditioned medium, the loss of biotinylated cell-surface HB-EGF(TM)HA/Myc, and
      the appearance of a Myc-tagged cytoplasmic tail fragment in cell lysates. By
      using this approach, several novel metalloproteinase-dependent regulators of
      HB-EGF(TM) shedding were identified as follows. (i) HB-EGF(TM)HA/Myc shedding
      induced by PMA was blocked by the mitogen-activated protein (MAP) kinase kinase
      inhibitor, PD98059. PMA activated MAP kinase within 5 min, but HB-EGF(TM)HA/Myc
      shedding did not occur until 20 min, suggesting that MAP kinase activation was a 
      necessary step in the pathway of PMA-induced HB-EGF(TM) cleavage. (ii) Activation
      of an inducible Raf-1 kinase, DeltaRaf-1:estrogen receptor, resulted in a rapid
      MAP kinase activation within 10 min and shedding of HB-EGF(TM)HA/Myc within 20-40
      min. (iii) Serum induced MAP kinase activation and HB-EGF(TM)HA/Myc shedding that
      were inhibited by PD98059. (iv) Whereas PMA induced HB-EGF(TM)HA/Myc shedding in 
      attached cells, no shedding occurred when the cells were placed in suspension.
      Shedding was fully restored shortly after cells were allowed to spread on
      fibronectin, and the extent of PMA-induced shedding increased with the extent of 
      cell spreading. PMA induced the same level of MAP kinase activation whether the
      cells were attached or in suspension suggesting that although MAP kinase
      activation might be necessary for shedding, it was not sufficient. Taken
      together, these results suggest that there are two components of cell regulation 
      that contribute to the shedding process, not previously recognized, the Raf-1/MAP
      kinase signal transduction pathway and cell adhesion and spreading.
FAU - Gechtman, Z
AU  - Gechtman Z
AD  - Department of Surgical Research, Children's Hospital Harvard Medical School,
      Boston, Massachusetts 02115, USA.
FAU - Alonso, J L
AU  - Alonso JL
FAU - Raab, G
AU  - Raab G
FAU - Ingber, D E
AU  - Ingber DE
FAU - Klagsbrun, M
AU  - Klagsbrun M
LA  - eng
GR  - CA 45548/CA/NCI NIH HHS/United States
GR  - GM RO1.47397/GM/NIGMS NIH HHS/United States
GR  - HL 56398/HL/NHLBI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (DNA Primers)
RN  - 0 (DNA, Complementary)
RN  - 0 (Heparin-binding EGF-like Growth Factor)
RN  - 0 (Intercellular Signaling Peptides and Proteins)
RN  - 0 (Retroviridae Proteins, Oncogenic)
RN  - 62229-50-9 (Epidermal Growth Factor)
RN  - EC 2.7.11.1 (Oncogene Proteins v-raf)
RN  - EC 3.4.24.- (Metalloendopeptidases)
RN  - NI40JAQ945 (Tetradecanoylphorbol Acetate)
SB  - IM
SB  - S
MH  - Animals
MH  - Base Sequence
MH  - Blood
MH  - CHO Cells
MH  - *Cell Adhesion
MH  - Cell Membrane/metabolism
MH  - *Cell Movement
MH  - Cricetinae
MH  - DNA Primers
MH  - DNA, Complementary
MH  - Epidermal Growth Factor/*metabolism
MH  - Heparin-binding EGF-like Growth Factor
MH  - Hydrolysis
MH  - Intercellular Signaling Peptides and Proteins
MH  - *MAP Kinase Signaling System
MH  - Metalloendopeptidases/metabolism
MH  - Oncogene Proteins v-raf
MH  - Retroviridae Proteins, Oncogenic/*metabolism
MH  - Tetradecanoylphorbol Acetate/pharmacology
OTO - NASA
OT  - Non-programmatic
EDAT- 1999/09/25 00:00
MHDA- 1999/09/25 00:01
CRDT- 1999/09/25 00:00
PHST- 1999/09/25 00:00 [pubmed]
PHST- 1999/09/25 00:01 [medline]
PHST- 1999/09/25 00:00 [entrez]
AID - 10.1074/jbc.274.40.28828 [doi]
PST - ppublish
SO  - J Biol Chem. 1999 Oct 1;274(40):28828-35. doi: 10.1074/jbc.274.40.28828.