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J. Biol. Chem., Vol. 276, Issue 20, 16772-16779, May 18, 2001
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From the Pleiotrophin (PTN) is a secreted growth factor
that induces neurite outgrowth and is mitogenic for fibroblasts,
epithelial, and endothelial cells. During tumor growth PTN can serve as
an angiogenic factor and drive tumor invasion and metastasis. To identify a receptor for PTN, we panned a phage display human cDNA library against immobilized PTN protein as a bait. From this we isolated a phage insert that was homologous to an amino acid sequence stretch in the extracellular domain (ECD) of the orphan receptor tyrosine kinase anaplastic lymphoma kinase (ALK). In parallel with PTN,
ALK is highly expressed during perinatal development of the nervous
system and down-modulated in the adult. Here we show in cell-free
assays as well as in radioligand receptor binding studies in intact
cells that PTN binds to the ALK ECD with an apparent
Kd of 32 ± 9 pM. This receptor
binding is inhibited by an excess of PTN, by the ALK ECD, and by
anti-PTN and anti-ECD antibodies. PTN added to ALK-expressing cells
induces phosphorylation of both ALK and of the downstream effector
molecules IRS-1, Shc, phospholipase C- The nucleotide sequences reported in this paper have been
submitted to the GenBankTM/EBI Data Bank with accession numbers
U66559, M57399, AF236106, and AF149800.
Identification of Anaplastic Lymphoma Kinase as a Receptor
for the Growth Factor Pleiotrophin*
§,§,,,,,
,, and**
Lombardi Cancer Center, Georgetown
University, Washington, D.C. 20007, ¶ AMGEN, Thousand Oaks,
California 91320, and Ciphergen Biosystems, 490 San Antonio
Road, Palo Alto, California 94306
, and phosphatidylinositol
3-kinase. Furthermore, the growth stimulatory effect of PTN on
different cell lines in culture coincides with the endogenous
expression of ALK mRNA, and the effect of PTN is enhanced by ALK
overexpression. From this we conclude that ALK is a receptor that
transduces PTN-mediated signals and propose that the PTN-ALK axis can
play a significant role during development and during disease processes.
*
This work was supported by National Institutes of Health
Grant CA58185 (Special Program of Research Excellence) and by a
United States Army Medical Research Materiel Command Breast
Cancer Program grant (to A. W.) as well as funds from the
Studienstiftung des Deutschen Volkes (to A. A.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
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