COMMUNICATION
Decorin Is a Biological Ligand for the Epidermal Growth
Factor Receptor*
Renato V.
Iozzo
§¶,
David K.
Moscatello§,
David J.
McQuillan
, and
Inge
Eichstetter
From the
Department of Pathology, Anatomy,
and Cell Biology and § Kimmel Cancer Center, Thomas
Jefferson University, Philadelphia, Pennsylvania 19107 and
Center for Extracellular Matrix Biology, Institute of
Biosciences and Technology, Texas A&M University,
Houston, Texas 77030
 |
ABSTRACT |
Ectopic expression of decorin induces profound
cytostatic effects in transformed cells with diverse histogenetic
backgrounds. The mechanism of action has only recently begun to be
elucidated. Exogenous decorin activates the epidermal growth factor
(EGF) receptor, thereby triggering a signaling cascade that leads to phosphorylation of mitogen-activated protein (MAP) kinase, induction of
p21, and growth suppression. In this study we demonstrate a direct
interaction of decorin with the EGF receptor. Binding of decorin
induces dimerization of the EGF receptor and rapid and sustained
phosphorylation of MAP kinase in squamous carcinoma cells. In a
cell-free system, decorin induces autophosphorylation of purified EGF
receptor by activating the receptor tyrosine kinase and can also
act as a substrate for the EGF receptor kinase itself. Using
radioligand binding assays we show that both immobilized and soluble
decorin bind to the EGF receptor ectodomain or to purified EGF
receptor. The binding is mediated by the protein core and has
relatively low affinity (Kd ~87 nM).
Thus, decorin should be considered as a novel biological ligand for the
EGF receptor, an interaction that could regulate cell growth during
remodeling and cancer growth.
 |
INTRODUCTION |
Decorin plays a pivotal role in regulating the proper assembly of
collagenous matrices and in the control of cell proliferation (1). Most
of its biological interactions occur via the central leucine-rich
repeat region, which is thought to fold into an arch-shaped structure
(2). Because of the ability of decorin to bind fibrillar collagen and
to delay in vitro fibrillogenesis (3), this proteoglycan is
regarded as a key modulator of matrix assembly. Genetic studies utilizing decorin null mice have indeed proved a major role for decorin
in the homeostasis of dermal collagen. Although the nullizygous animals
grew to adulthood without any overt pathology, a close analysis
revealed a skin fragility phenotype; the dermal collagen exhibited
aberrant organization of fibrils with abnormal packing and a great
variability in diameter (4). Another important role of decorin is
linked to its ability to inhibit transforming growth factor-
, which
in turns blocks cell proliferation (5).
There is also mounting evidence that decorin is involved directly in
the control of cell growth. Decorin levels are markedly elevated during
growth arrest and quiescence, its expression is abrogated by viral
transformation, and its transcription is suppressed in most tumorigenic
cell lines (6). De novo expression of decorin induces a
generalized growth suppression by activating p21, a potent inhibitor of
cyclin-dependent kinase activity (7-9). Decorin activates
the EGF1 receptor (EGFR) (10)
that in turn triggers a signal cascade leading to activation of MAP
kinases, mobilization of intracellular calcium (11), up-regulation of
p21, and growth suppression.
In this report we demonstrate a direct interaction between the
EGFR and decorin. Exogenous decorin causes dimerization of the EGFR in
A431 squamous carcinoma cells. Decorin induces autophosphorylation of
the EGFR by activating the EGFR tyrosine kinase in a cell-free system
and can also act as a substrate for the EGFR kinase itself. Both
immobilized and soluble decorin bind to the soluble EGFR ectodomain or
to immunopurified EGFR. The binding is solely mediated by the protein
core and has relatively low affinity (Kd ~87
nM). Thus, decorin should be considered as a novel
biological ligand for the EGFR.
 |
EXPERIMENTAL PROCEDURES |
Materials--
Media and fetal bovine serum were obtained from
Hyclone Laboratories, Inc. (Logan, UT). [
-32P]ATP
(6000 Ci/mmol) and Hybond ECL membranes were from Amersham Pharmacia
Biotech. The EGFR preparation purified from A431 cells by
immunoaffinity chromatography (Sigma) contained ~500 units of active
EGFR; 1 unit catalyzes the incorporation of 1 pmol of phosphate/min
from [
-32P]ATP into poly(Glu,Tyr) at 30 °C.
Cross-linking Studies and Activation of MAP
Kinases--
Confluent A431 cells, which were rendered quiescent by
serum deprivation for 36 h, were incubated with decorin or decorin protein core (1 µM) or EGF (16 nM) or various
combinations at 4 °C for 1 h. The cells were washed extensively
and incubated for 10 min with the membrane-impermeable cross-linker
bis[sulfosuccinimidyl]suberate (BS3, (Pierce). The cell
lysates were separated in 3-15% SDS-PAGE and subjected to
immunoblotting (10) with anti-EGFR antibody. The anti-phospho-MAP
kinase antibody (New England BioLabs) detects p42 and p44 MAP kinases
(Erk1/Erk2) only when they are catalytically activated by
phosphorylation at Thr202 and Tyr204.
In Vitro Phosphorylation of the EGFR Kinase--
Constant
amounts (~300 ng) of immunopurified EGFR were preincubated with
buffer alone or containing increasing concentrations (5-20 µg) of
decorin, its decorin protein core, or collagen type I. After 15 min of
incubation in kinase buffer (20 mM HEPES, pH 7.4, 2 mM MnCl2, 10 mM
p-nitrophenyl phosphate, 40 µM
Na3VO4, 0.01% BSA, 15 µM ATP) 1 µCi of [
-32P]ATP and 0.2% Nonidet P-40 was added to
reach a final volume of 60 µl. The mixture was incubated for an
additional 10 min, stopped by boiling in SDS buffer, and analyzed by
SDS-PAGE. Phosphorylated proteins were visualized by autoradiography.
Control samples omitting either [
-32P]ATP or EGFR
showed no activity.
Binding Studies with Soluble EGFR Ectodomain--
Because A431
cells synthesize a soluble form of EGFR of ~105 kDa lacking the
transmembrane and intracytoplasmic domains (12), serum-free medium
conditioned by confluent A431 cells was concentrated by Centricon-50
and incubated with decorin, or its protein core immobilized on
nitrocellulose filters, washed, and subjected to immunodetection with
anti-EGFR antiserum. Briefly, serial dilutions of BSA, decorin, or its
protein core were slot-blotted onto ECL nitrocellulose, blocked
overnight at 4 °C with 5% fetal bovine serum and 5% nonfat milk,
and washed several times in TBS-T (Tris-buffered saline, 25 mM Tris-HCl, 150 mM NaCl, 0.1% Tween 20, pH
7.4). The membranes were incubated with the serum-free medium
conditioned by the A431 cells, washed 3 more times, incubated with an
antibody raised against the N-terminal sequence LEEKK of the human
EGFR, and affinity-purified on a peptide linked to Sepharose. This
antibody would not interfere with the binding because it recognizes the N-terminal end of the EGFR. As additional control, we used a mouse monoclonal antibody 225 raised against the EGF-binding domain of the
EGFR (13).
Interaction Between Decorin and Purified EGF Receptor--
We
utilized columns containing nickel-nitrilotriacetic acid (Ni-NTA) as
ligand (Qiagen) since the recombinant human decorin contains an
N-terminal tag with 6 histidine residues (His6) that allows
a rapid and efficient purification via Ni-NTA affinity chromatography
(14). First we phosphorylated the immunopurified EGFR to reach a
specific activity of ~1.7 × 1017 cpm/mol using the
in vitro phosphorylation assay described above. Following
purification by Sephadex G-50 chromatography, constant amounts of
32P-labeled EGFR were incubated with increasing
concentrations of decorin or its core protein for 30 min at 4 °C
under gentle agitation. The spin columns were equilibrated with 3 column volumes of binding buffer (300 mM
NaH2PO4, pH 8.0, 300 mM NaCl), and
then the samples were applied and spun at 750 × g for
5 min. Following two consecutive washes, the bound decorin-EGFR
complexes were eluted with buffer containing 250 mM
imidazole. Parts of the fractions were counted in a scintillation
counter, and parts were analyzed by SDS-PAGE and autoradiography.
Radioligand binding assays were performed as described before (15).
Decorin was coated in removable Immulon 4HXB wells (Dynex Technologies,
Chantilly, VA), and binding of 32P-labeled EGFR was
measured in TBS supplemented with 2 mM CaCl2, 2 mM MgCl2, 0.02% NaN3, and 1 mg/ml
heat-inactivated BSA following incubation under gentle shaking (60 rpm)
for 4-14 h. Reversible binding was demonstrated by incubation with
100-fold molar excess EGFR. After incubation the wells were washed with
ice-cold TBS-T and then counted in toto. Scatchard plots
were generated using the Ligand program as described before (16).
 |
RESULTS AND DISCUSSION |
Decorin Causes Dimerization of the EGF Receptor--
It is well
established that the general mechanism transmitting extracellular
signals starts with binding of a growth factor to a cell surface
receptor that in many cases carries an intrinsic tyrosine kinase
activity (17). For example, EGF stimulates the formation of
non-covalent homo- and heterodimers with other members of the family of
receptor tyrosine kinase (17). The growth of some tumor cells bearing
high levels of EGF receptors is paradoxically suppressed. For instance,
A431 vulvar squamous carcinoma cells are stimulated to grow with
picomolar amounts of EGF but are markedly growth inhibited with
nanomolar amounts of the growth factor (18). EGF treatment of A431
cells causes a protracted induction of p21(19, 20), and similarly,
antibodies that recognize the EGF-binding region of the EGFR inhibit
the proliferation of normal and transformed cells (13, 21-24). Thus,
we reasoned that decorin might induce growth suppression by activating
a similar signal-transducing pathway. To test whether decorin could
cause dimerization of the EGFR, we added purified decorin proteoglycan
or its protein core to quiescent A431 cells at 4 °C for 1 h,
washed extensively, and incubated for 10 min with BS3, a
non-cleavable, membrane-impermeable cross-linker. Decorin induced
significant dimerization of the EGFR (Fig.
1A, lanes
4 and 8), estimated to be ~40% of that induced
by EGF (Fig. 1A, lane 2). There were
no additive effects when decorin or its protein core were added
concurrently with EGF (Fig. 1, A, lanes
6 and 10, and B, lane
6). The protein core of decorin was capable of mediating the
full effect of the decorin proteoglycan (Fig. 1B). To
investigate this further, we iodinated decorin and performed similar
cross-linking experiments followed by immunoprecipitation. We found a
very high Mr complex that could not penetrate
the gel (not shown), further suggesting the decorin might be directly involved in binding to the EGFR.

View larger version (46K):
[in this window]
[in a new window]
|
Fig. 1.
Decorin causes dimerization of the EGF
receptor in A431 cells. Confluent A431 cells were incubated with
decorin (Dcn), decorin protein core (1 µM),
EGF (16 nM), or the various combinations as indicated at
4 °C for 1 h. The cells were washed extensively and incubated
for 10 min with the membrane-impermeable cross-linker BS3
(Pierce). The cell lysates were separated in 3-15% SDS-PAGE and
subjected to Western immunoblotting (10) with anti-EGFR antibody. The
migration of the EGFR dimer (~340 kDa) and monomer (~170 kDa) is
indicated on the right.
|
|
Decorin Causes Rapid and Sustained Activation of MAP Kinase and
Induces Autophosphorylation of Purified EGF Receptor--
To
investigate whether decorin induces rapid and sustained activation of
the MAP kinase signal-transducing pathway, we exposed quiescent A431
cells to 1 µM decorin or its protein core for either 18 h or 10 min. As positive and negative controls we used EGF (16 nM) and collagen (1 µM), respectively. The
results showed both a sustained (Fig.
2A) and rapid (Fig. 2,
B and C) activation of the MAP kinase signal
pathway, which leads to prolonged activation of the endogenous p21 and
block of the cells in G1 (10). To more directly investigate
the interaction between the EGFR and decorin, we used EGFR purified by
immunoaffinity chromatography from plasma membranes of A431 cells (25).
The EGFR was not exposed to EGF during the purification procedure, and
thus it contains optimal kinase activity and is suitable for both
binding and in vitro kinase assays. First, we established by
Western immunoblotting the purity of the EGFR and the lack of
degradation products. A single ~170-kDa band was seen using an
antibody against the N terminus of the EGFR (Fig. 2D). When
the EGFR was subjected to in vitro kinase assay there was a
dose-dependent induction of EGFR autophosphorylation only
in the presence of decorin (Fig. 2E, lanes
2-4) or its protein core (Fig. 2E,
lanes 5-7) but none in the presence of collagen
(Fig. 2E, lanes 8-10). Of note, the EGFR kinase efficiently phosphorylated decorin protein core as seen by
the appearance of the two ~42- and 46-kDa proteins seen only when the
protein core was added (Fig. 2E, lanes
5-7). No significant phosphorylation of the decorin
proteoglycan (which would have appeared as a ~100-kDa band) was
noted, indicating that the actual phosphorylation of the protein core
may be inhibited by the dermatan sulfate chain. The binding of decorin
to the EGFR was totally abolished by thermal denaturation (80 °C for
15 min) of both decorin and its protein core (not shown). The same
treatment of the EGFR abolished the binding to decorin. Thus, proper
protein folding is required for this interaction.

View larger version (41K):
[in this window]
[in a new window]
|
Fig. 2.
Decorin activates MAP kinases in A431 cells
and induces autophosphorylation of purified EGF receptor.
A-C, Western immunoblotting of total cell lysate probed
with an antibody against the phosphorylated forms of Erk1/Erk2 MAP
kinases (MAPK) using a polyclonal anti- phospho-p44/42 MAP
kinase antibody (New England BioLabs). Cells were rendered quiescent by
serum starvation for 2 days and then treated with 1 µM
decorin, protein core, or collagen type I as indicated. EGF (~16
nM) was used as a positive control (10). D,
Western immunoblot of immunopurified EGFR using an antiserum against
the N-terminal domain of the EGFR. E, in vitro
phosphorylation assay using immunopurified EGFR and decorin, its
protein core, or collagen. Constant amounts (~300 ng) of
immunopurified EGFR were preincubated with buffer alone
(lane 1) or containing decorin (lanes
2-4; at 5, 10, and 20 µg, respectively), decorin protein
core (lanes 5-7; at 5, 10, and 20 µg,
respectively), or collagen (lanes 8-10; at 5, 10, and 20 µg, respectively). After 15 min in kinase buffer, 1 µCi
of [ -32P]ATP and 0.2% Nonidet P-40 was added. The
mixture was incubated for an additional 10 min, stopped by boiling in
SDS buffer, and analyzed by SDS-PAGE and autoradiography.
|
|
Decorin Interacts with the Soluble Ectodomain of the EGF
Receptor--
The rationale for these studies is based on the
observation that A431 cells synthesize and release into the medium a
soluble form of EGFR of ~105 kDa lacking the transmembrane and
intracytoplasmic domains (12, 26). This soluble protein is derived from
the 2.8-kilobase mRNA transcribed from a rearranged EGFR gene on
chromosome 7 (27). As the extracellular domain has both a high Cys
content and large amount of oligosaccharides, the ectodomain is quite stable and highly resistant to proteolysis. It binds with high affinity
EGF (12, 26), and available stoichiometry data show that 1 mol of EGFR
binds 1 mol of EGF. First, we tested the purity of the medium
conditioned for 48 h by A431 cells by performing Western
immunoblotting. A single band of ~105 kDa was detected using the
anti-EGFR antiserum (Fig. 3A).
This material was used in slot-blot overlay assays in which both
proteins (the immobilized and the soluble ligand) are under native
conditions. Serum-free medium conditioned for 48 h by A431 cells
was concentrated ~8-fold by Centricon-50 and incubated with decorin,
its protein core, or BSA immobilized on nitrocellulose filters, washed,
and subjected to immunodetection with anti-EGFR antiserum (28). Both
protein core (Fig. 3B) and decorin (Fig. 3C)
bound specifically to soluble EGFR ectodomain, in contrast to BSA,
which was unreactive. Of note, a chimeric biglycan/decorin protein
containing the N terminus of biglycan and the remaining decorin gave
identical results (not shown). Thus, the binding to EGFR is not
mediated by the N terminus since biglycan did not cause activation of
EGFR (10). When similar slot-blot overlay assays were tested with
monoclonal antibody 225 that reacts specifically with the EGF binding
region of the EGFR (13), there was no reactivity (not shown),
suggesting that decorin might bind near or perhaps in the same
location where EGF binds.

View larger version (35K):
[in this window]
[in a new window]
|
Fig. 3.
Immobilized decorin or its protein core
specifically interact with the soluble ectodomain of the EGF
receptor. A, Western immunoblotting of serum-free media
conditioned for 48 h by A431 cells. A single band of ~105 kDa
corresponding to the soluble EGFR ectodomain was detected using the
anti-EGFR antiserum. B and C, slot-blot overlay
assays showing specific binding of the EGFR ectodomain to decorin or
its protein core. Serum-free media conditioned for 48 h by A431
cells was concentrated ~8-fold and incubated with scalar dilutions
(as indicated in the right margins) of decorin,
its protein core, or BSA immobilized on nitrocellulose filters. The
filters were washed and subjected to immunodetection with anti-EGFR
antiserum (28). DMEM, Dulbecco's modified Eagle's medium.
D, interaction of soluble decorin with immobilized media
conditioned by A431 cells for 24 or 48 h or with medium alone.
Aliquots (25-400 µl) were applied by vacuum to Hybond ECL
nitrocellulose membranes, blocked overnight at 4 °C in TBS-T
containing 1.5% BSA, washed three times, and then incubated for 1 h in 1 µg/ml decorin (~10 pmol). The membranes were washed as
before and incubated for 1 h with LF-30 rabbit anti-human decorin
antibody (1:1000) and detected by ECL. E, scalar dilutions
of decorin served as positive control for the overlay assay with
anti-decorin antibody.
|
|
To further corroborate the above results, we immobilized on
nitrocellulose scalar amounts of media conditioned for various periods
of time by A431 cells and performed overlay assays with soluble decorin
followed by Western immunoblotting (29). The results showed specific
binding of decorin to serum-free medium conditioned by A431 cells,
which contains the EGFR ectodomain (Fig. 3D). As negative
and positive controls we used either Dulbecco's modified Eagle's
medium (Fig. 3D) or scalar dilutions (50-0 ng) of purified
decorin (Fig. 3E), respectively. Our results show that
decorin interacts specifically with the ectodomain of the EGFR either
in solution or immobilized on nitrocellulose membranes.
Decorin Interacts with Purified EGF Receptor--
Next we
sought to investigate whether soluble decorin could interact with the
EGFR under physiological salt concentrations and in solution. The
rationale for this approach is that decorin contains an N-terminal
His6 tag that allows a rapid and efficient purification via
Ni-NTA affinity chromatography (14). Constant amounts of
32P-labeled EGFR were incubated with increasing
concentrations of decorin or its core protein for 30 min at 4 °C
under gentle agitation. The Ni-NTA columns were equilibrated with 3 column volumes of binding buffer, and then the samples were applied and
spun at 750 × g for 5 min. Following two consecutive
washes, the bound decorin-EGFR complexes were eluted with buffer
containing 250 mM imidazole. In two independent experiments
there was specific binding of the EGFR to decorin since the amount of
radioactive EGFR increased proportionally to the amount of interacting
(Ni-NTA-bound) decorin (Fig. 4,
A-C). Quantitation of both experiments gave a strong
correlation with r2 = 0.961 and
p < 0.0005 (Fig. 4D).

View larger version (30K):
[in this window]
[in a new window]
|
Fig. 4.
Decorin specifically interacts with EGF
receptor. A and B, mean values of two
independent experiments showing the amount of radiolabeled EGFR eluted
with 250 mM imidazole versus increasing
concentrations of decorin protein core. The radiolabeled EGFR interacts
specifically with decorin, which in turn binds with high affinity to
the Ni-NTA column via the polyhistidine tag. C,
autoradiogram of 32P-labeled EGFR that was bound to the
Ni-NTA column. The numbers at the bottom
correspond to the same concentrations of decorin protein core shown in
A. D, relationship between EGFR and increasing
amounts of decorin protein core. Linear regression, correlation
coefficient (r2), and p values were
obtained using the Regression Wizard Program of the Sigma Plot 4 Package (Jandel). Saturation curve (E) and Scatchard plot
(F) for the binding or 32P-labeled EGFR to
decorin protein core are shown. The EGFR was tested at 0.1-3.2 pmol
(1.7 × 1017 cpm/mol) on removable wells coated with 1 µg (~22 pmol) of decorin protein core. The values represent the
average of two independent experiments. The cumulative data from five
experiments gave a Kd ~87 nM ± 7.5.
|
|
Finally, to determine more precisely the binding affinity we utilized a
radioligand binding assay where increasing concentrations of
32P-labeled EGFR were tested on Immulon wells coated with
decorin protein core (22 pmol). Binding of EGFR to decorin protein core was saturable (Fig. 4E), and a Scatchard plot (Fig.
4F) gave a single straight line with Kd
~87 ± 7.5 nM (n = 5). The binding
of decorin to the radiolabeled EGFR was totally abolished by 100-fold
molar excess of cold EGFR (not shown). In parallel experiments where
10-fold more decorin protein core was used to coat the wells, the
binding of EGFR was rapidly saturable, and the data also yielded linear
Scatchard plots (not shown). Notably, the affinity constant obtained in
such experiments is ~90-fold lower than that reported for the low
affinity binding sites for EGF (~1 nM) and ~900-fold
lower than the high affinity receptor for EGF (~0.1 nM)
(27).
Conclusions--
We have demonstrated that decorin protein core
causes a rapid phosphorylation of the EGFR in A431 cells, which leads
to a specific activation of the MAP kinase signal pathway and to
induction of endogenous p21. Several lines of evidence support a
specific protein/protein interaction between decorin and the EGFR. We
demonstrate for the first time that decorin is capable of inducing
dimerization of the EGFR in live cells, a physiologic phenomenon
previously shown to be a prelude to receptor activation. The specific
binding occurs when decorin is either immobilized on nitrocellulose
membranes or free in physiologic salt solutions. In a cell-free system, decorin induces autophosphorylation of purified EGFR by activating the
receptor tyrosine kinase and can also act as a substrate for the EGFR
kinase itself, although it is unlikely to be a substrate in
vivo since decorin is an extracellular molecule. Decorin is capable of inducing EGFR tyrosine kinase and that both the binding and
activation require properly folded protein moiety. Notably, DDR1 and
DDR2, two "orphan" receptor tyrosine kinases, have been shown to be
the receptors for fibrillar collagen (28, 30). Similarly to the
decorin/EGFR interaction, stimulation of the DDR receptor tyrosine
kinase activity requires the native triple helical structure of
collagen and occurs over an extended period of time (30). The
unexpected realization that extracellular matrix molecules can directly
serve as ligands for receptor tyrosine kinases may change prevailing
views about the mechanisms by which cells perceive and respond to the
extracellular matrix signals. Because decorin is intimately associated
with fibrillar collagen, a complex scenario where multimeric
interactions take place in an integrin-independent manner should be
considered. An increase in decorin content in the newly formed tumor
stroma could potentially trigger a functional interaction with the
EGFR, known to be highly expressed in most tumor cells, which would in
turn start a signaling process that directly influences the cell cycle machinery.
 |
ACKNOWLEDGEMENTS |
We thank L. Fisher, A. Wong,
and Z. Fan for providing antibodies and K. G. Danielson for help
in the initial stages of this project.
 |
FOOTNOTES |
*
This work was supported by National Institutes of Health
Grants RO1 CA39481 and RO1 CA47282 (to R. V. I.) and RO1 AR42826 (to
D. J. M.).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.
This paper is dedicated to the memory of our beloved colleague and
friend, Renée K. Margolis.
¶
To whom correspondence should be addressed: Dept. of
Pathology, Anatomy, and Cell Biology, Rm. 249, JAH, Thomas Jefferson University, 1020 Locust St., Philadelphia, PA 19107. E-mail:
iozzo{at}lac.jci.tju.edu.
 |
ABBREVIATIONS |
The abbreviations used are:
EGF, epidermal
growth factor;
EGFR, EGF receptor;
p21, the
cyclin-dependent kinase inhibitor p21WAF1/CIP1;
BSA, bovine serum albumin;
MAP, mitogen-activated protein;
PAGE, polyacrylamide gel electrophoresis;
NTA, nitrilotriacetic acid;
BS3, bis[sulfosuccinimidyl]suberate.
 |
REFERENCES |
-
Iozzo, R. V.
(1998)
Annu. Rev. Biochem.
67,
609-652[CrossRef][Medline]
[Order article via Infotrieve]
-
Weber, I. T.,
Harrison, R. W.,
and Iozzo, R. V.
(1996)
J. Biol. Chem.
271,
31767-31770[Abstract/Free Full Text]
-
Vogel, K. G.,
Paulsson, M.,
and Heinegård, D.
(1984)
Biochem. J.
223,
587-597[Medline]
[Order article via Infotrieve]
-
Danielson, K. G.,
Baribault, H.,
Holmes, D. F.,
Graham, H.,
Kadler, K. E.,
and Iozzo, R. V.
(1997)
J. Cell Biol.
136,
729-743[Abstract/Free Full Text]
-
Border, W. A.,
Noble, N. A.,
Yamamoto, T.,
Harper, J. R.,
Yamaguchi, Y.,
Pierschbacher, M. D.,
and Ruoslahti, E.
(1992)
Nature
360,
361-364[CrossRef][Medline]
[Order article via Infotrieve]
-
Mauviel, A.,
Santra, M.,
Chen, Y. Q.,
Uitto, J.,
and Iozzo, R. V.
(1995)
J. Biol. Chem.
270,
11692-11700[Abstract/Free Full Text]
-
Santra, M.,
Skorski, T.,
Calabretta, B.,
Lattime, E. C.,
and Iozzo, R. V.
(1995)
Proc. Natl. Acad. Sci. U. S. A.
92,
7016-7020[Abstract]
-
De Luca, A.,
Santra, M.,
Baldi, A.,
Giordano, A.,
and Iozzo, R. V.
(1996)
J. Biol. Chem.
271,
18961-18965[Abstract/Free Full Text]
-
Santra, M.,
Mann, D. M.,
Mercer, E. W.,
Skorski, T.,
Calabretta, B.,
and Iozzo, R. V.
(1997)
J. Clin. Invest.
100,
149-157[Abstract/Free Full Text]
-
Moscatello, D. K.,
Santra, M.,
Mann, D. M.,
McQuillan, D. J.,
Wong, A. J.,
and Iozzo, R. V.
(1998)
J. Clin. Invest.
101,
406-412[Abstract/Free Full Text]
-
Patel, S.,
Santra, M.,
McQuillan, D. J.,
Iozzo, R. V.,
and Thomas, A. P.
(1998)
J. Biol. Chem.
273,
3121-3124[Abstract/Free Full Text]
-
Weber, W.,
Gill, G. N.,
and Spiess, J.
(1984)
Science
224,
294-297[Medline]
[Order article via Infotrieve]
-
Fan, Z.,
Lu, Y.,
Wu, X.,
and Mendelsohn, J.
(1994)
J. Biol. Chem.
269,
27595-27602[Abstract/Free Full Text]
-
Ramamurthy, P.,
Hocking, A. M.,
and McQuillan, D. J.
(1996)
J. Biol. Chem.
271,
19578-19584[Abstract/Free Full Text]
-
Milev, P.,
Chiba, A.,
Margolis, R. K.,
Schachner, M.,
Ranscht, B.,
and Margolis, R. U.
(1998)
J. Biol. Chem.
273,
6998-7005[Abstract/Free Full Text]
-
Milev, P.,
Monnerie, H.,
Popp, S.,
Margolis, R. K.,
and Margolis, R. U.
(1998)
J. Biol. Chem.
273,
21439-21442[Abstract/Free Full Text]
-
van der Geer, P.,
Hunter, T.,
and Lindberg, R. A.
(1994)
Annu. Rev. Cell Biol.
10,
251-337[CrossRef]
-
Barnes, D. W.
(1982)
J. Cell Biol.
93,
1-4[Abstract]
-
Fan, Z.,
Lu, Y.,
Wu, X.,
DeBlasio, A.,
Koff, A.,
and Mendelsohn, J.
(1995)
J. Cell Biol.
131,
235-242[Abstract]
-
Jakus, J.,
and Yeudall, W. A.
(1996)
Oncogene
12,
2369-2376[Medline]
[Order article via Infotrieve]
-
Fan, Z.,
Baselga, J.,
Masui, H.,
and Mendelson, J.
(1993)
Cancer Res.
53,
4637-4642[Abstract]
-
Fan, Z.,
Mendelsohn, J.,
Masui, H.,
and Kumar, R.
(1993)
J. Biol. Chem.
268,
21073-21079[Abstract/Free Full Text]
-
Van de Vijver, M.,
Kumar, R.,
and Mendelsohn, J.
(1991)
J. Biol. Chem.
266,
7503-7508[Abstract/Free Full Text]
-
Fan, Z.,
Masui, H.,
Altas, I.,
and Mendelsohn, J.
(1993)
Cancer Res.
53,
4322-4328[Abstract]
-
Yarden, Y.,
Harari, I.,
and Schlessinger, J.
(1985)
J. Biol. Chem.
260,
315-319[Abstract/Free Full Text]
-
Weber, W.,
Bertics, P. J.,
and Gill, G. N.
(1984)
J. Biol. Chem.
259,
14631-14636[Abstract/Free Full Text]
-
Carpenter, G.
(1987)
Annu. Rev. Biochem.
56,
881-914[CrossRef][Medline]
[Order article via Infotrieve]
-
Shrivastava, A.,
Radziejewski, C.,
Campbell, E.,
Kovac, L.,
McGlynn, M.,
Ryan, T. E.,
Davis, S.,
Goldfarb, M. P.,
Glass, D. J.,
Lemke, G.,
and Yancopoulos, G. D.
(1997)
Mol. Cell
1,
25-34[Medline]
[Order article via Infotrieve]
-
Fisher, L. W.,
Stubbs, J. T., III,
and Young, M. F.
(1995)
Acta Orthop. Scand.
66,
61-65
-
Vogel, W.,
Gish, G. D.,
Alves, F.,
and Pawson, T.
(1997)
Mol. Cell
1,
13-23[Medline]
[Order article via Infotrieve]
Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.