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J Biol Chem, Vol. 274, Issue 18, 12376-12382, April 30, 1999
From the Platelet factor 4 (PF-4) is a platelet-derived
Characterization of a Neutrophil Cell Surface Glycosaminoglycan
That Mediates Binding of Platelet Factor 4
,
, and
Department of Medical Biochemistry and
Microbiology, Uppsala University, Biomedical Center, Box 575, S-75123 Uppsala, Sweden and the ¶ Department of Immunology and
Cell Biology, Research Center Borstel, Parkallee 22, D-23845 Borstel, Germany
-chemokine that binds to and activates human neutrophils to undergo
specific functions like exocytosis or adhesion. PF-4 binding has been
shown to be independent of interleukin-8 receptors and could be
inhibited by soluble chondroitin sulfate type glycosaminoglycans or by
pretreatment of cells with chondroitinase ABC. Here we present evidence
that surface-expressed neutrophil glycosaminoglycans are of chondroitin sulfate type and that this species binds to the tetrameric form of
PF-4. The glycosaminoglycans consist of a single type of chain with an
average molecular mass of ~23 kDa and are composed of ~85-90%
chondroitin 4-sulfate disaccharide units type CSA
(
4GlcA
1
3GalNAc(4-O-sulfate)
1
) and of
~10-15% di-O-sulfated disaccharide units. A major
part of these di-O-sulfated disaccharide units are CSE
units (
4GlcA
1
3GalNAc(4,6-O-sulfate)
1
). Binding studies revealed that the interaction of chondroitin sulfate with PF-4 required at least 20 monosaccharide units for significant binding. The di-O-sulfated disaccharide units in neutrophil
glycosaminoglycans clearly promoted the affinity to PF-4, which showed
a Kd ~ 0.8 µM, as the affinities of
bovine cartilage chondroitin sulfate A, porcine skin dermatan sulfate,
or bovine cartilage chondroitin sulfate C, all consisting exclusively
of monosulfated disaccharide units, were found to be 3-5-fold lower.
Taken together, our data indicate that chondroitin sulfate chains
function as physiologically relevant binding sites for PF-4 on
neutrophils and that the affinity of these chains for PF-4 is
controlled by their degree of sulfation.
Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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