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J Biol Chem, Vol. 275, Issue 6, 4374-4382, February 11, 2000
From the Secreted Frizzled-related protein-1 (sFRP-1)
contains a cysteine-rich domain homologous to the putative Wnt-binding
site of Frizzleds. To facilitate the biochemical and biological
analysis of sFRP-1, we developed a mammalian recombinant expression
system that yields ~3 mg of purified protein/liter of conditioned
medium. Using this recombinant protein, we demonstrated that sFRP-1 and Wg (wingless) interact in enzyme-linked immunosorbent and
co-precipitation assays. Surprisingly, a derivative lacking the
cysteine-rich domain retained the ability to bind Wg. Cross-linking
experiments performed with radioiodinated sFRP-1 provided definitive
evidence that sFRP-1 and Wg bind directly to each other. Besides
detecting a cross-linked complex consistent in size with 1:1
stoichiometry of sFRP-1 and Wg, we also observed a larger complex whose
size suggested the presence of a second sFRP-1 molecule. The formation
of both complexes was markedly enhanced by an optimal concentration of
exogenous heparin, emphasizing the potential importance of
heparan-sulfate proteoglycan in Wnt binding and signaling. sFRP-1
exerted a biphasic effect on Wg activity in an armadillo stabilization
assay, increasing armadillo level at low concentrations but reducing it
at higher concentrations. These results provide new insights about the
Wnt binding and biological activity of sFRPs.
Secreted Frizzled-related Protein-1 Binds Directly to Wingless
and Is a Biphasic Modulator of Wnt Signaling*
,,,,,¶
Laboratory of Cellular and Molecular
Biology, Division of Basic Sciences, NCI, National Institutes of
Health, Bethesda, Maryland 20892 and the § Department of
Biochemistry and Molecular Biology, University of Massachusetts,
Amherst, Massachusetts 01002
*
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.
Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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