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J Biol Chem, Vol. 274, Issue 12, 7741-7747, March 19, 1999
From the Heparin affin regulatory peptide (HARP) is a
polypeptide belonging to a family of heparin binding
growth/differentiation factors. The high affinity of HARP for heparin
suggests that this secreted polypeptide should also bind to heparan
sulfate proteoglycans derived from cell surface and extracellular
matrix defined as extracellular compartments. Using Western blot
analysis, we detected HARP bound to heparan sulfate proteoglycans in
the extracellular compartments of MDA-MB 231 and MC 3T3-E1 as well as
NIH3T3 cells overexpressing HARP protein. Heparitinase treatment of BEL
cells inhibited HARP-induced cell proliferation, and the biological activity of HARP in this system was restored by the addition of heparin. We report that heparan sulfate, dermatan sulfate, and to a
lesser extent, chondroitin sulfate A, displaced HARP bound to the
extracellular compartment. Binding analyses with a biosensor showed
that HARP bound heparin with fast association and dissociation kinetics
(kass = 1.6 × 106
M
Glycosaminoglycans Differentially Bind HARP and Modulate Its
Biological Activity
,,,,
Laboratoire de Recherche sur la Croissance
Cellulaire, la Réparation et la Régénération
Tissulaires (CRRET), Unité Propre de Recherche de l'Enseignement
Supérieur Associées an CNRS (UPRES-A) CNRS 7053, Université Paris XII-Val de Marne, avenue du
Général de Gaulle, 94010 Créteil, France and the
¶ School of Biological Sciences, University of Liverpool,
Liverpool L69 7ZB, United Kingdom
1 s1;
kdiss = 0.02 s1), yielding a
Kd value of 13 nM; the interaction
between HARP and dermatan sulfate was characterized by slower
association kinetics (kass = 0.68 × 106 M1 s1) and a
lower affinity (Kd = 51 nM). Exogenous
heparin, heparan sulfate, and dermatan sulfate potentiated the
growth-stimulatory activity of HARP, suggesting that corresponding
proteoglycans could be involved in the regulation of the mitogenic
activity of HARP.
Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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