Gsv. Kuschert et al., Glycosaminoglycans interact selectively with chemokines and modulate receptor binding and cellular responses, BIOCHEM, 38(39), 1999, pp. 12959-12968
Chemokines selectively recruit and activate a variety of cells during infla
mmation. Interactions between cell surface glycosaminoglycans (GAGs) and ch
emokines drive the formation of haptotactic or immobilized gradients of che
mokines at the site of inflammation, directing this recruitment. Chemokines
bind to glycosaminoglycans on human umbilical vein endothelial cells (HUVE
Cs) with affinities in the micromolar range: RANTES > MCP-1 > IL-8 > MIP-1
alpha. This binding can be competed with by soluble glycosaminoglycans: hep
arin, heparin sulfate, chondroitin sulfate, and dermatan sulfate. RANTES bi
nding showed the widest discrimination between glycosaminoglycans (700-fold
), whereas MIP-1 alpha was the least selective. Almost identical results we
re obtained in an assay using heparin sulfate beads as the source of immobi
lized glycosaminoglycan. The binding of chemokines to glycosaminoglycan fra
gments has a strong: length dependence, and optimally requires both N- and
O-sulfation. Isothermal titration calorimetry data confirm these results; I
L-8 binds heparin fragments with a K-d of 0.39-2.63 mu M, and requires five
saccharide units to bind each monomer of chemokine. In membranes from cell
s expressing the G-protein-coupled chemokine receptors CXCR1, CXCR2, and CC
R1, soluble GAGs inhibit the binding of chemokine ligands to their receptor
s, Consistent with this, heparin and heparin sulfate could inhibit IL-8-ind
uced neutrophil calcium flux. Chemokines can therefore form complexes with
both cell surface and soluble GAGs; these interactions have different funct
ions. Soluble GAG chemokines complexes are unable to bind the receptor, res
ulting in a block of the biological activity. Previously, we have shown tha
t cell surface GAGs present chemokines to the G-protein-coupled receptors,
by increasing the local concentration of protein. A model is presented whic
h brings together all of these data. The selectivity in the chemokine-GAG i
nteraction suggests selective disruption of the haptotactic gradient may be
an achievable therapeutic approach in inflammatory disease.