| Summary: | CD44 is a widely expressed cell adhesion molecule that binds the extracellular matrix
component, hyaluronan in a tightly regulated manner. As a molecule involved in adhesion and cell
migration, CD44 has been shown to interact with elements of the cytoskeleton via its cytoplasmic
domain. A significant portion of CD44 is Triton X-100 insoluble in fibroblasts and this has been
interpreted to reflect associations with the actin cytoskeleton. However, the detergent insolubility
of CD44 in NIH 3T3 cells cannot be attributed to an interaction with the cytoskeleton since CD44
lacking a cytoplasmic tail remains insoluble after Triton X-100 extraction. Instead, after
equilibrium density gradient centrifugation on sucrose, a proportion of CD44 was found in the low
density lipid fractions, indicating that CD44 insolubility in NIH 3T3 cells is due to an association
with Triton X-100-insoluble lipids. Furthermore, the migration of CD44 to the low density
fraction is dependent on the transmembrane domain of CD44, not the cytoplasmic domain.
Previous studies have indicated that the CD44-hyaluronan interaction is affected by changes
in the glycosylation state of CD44. Murine L cell variants defective in glycosaminoglycan
synthesis and oligosaccharide processing were used to assess the effects of these modifications on
CD44-mediated hyaluronan binding. Analysis of constitutive and antibody-induced hyaluronan
binding ability in these cells indicated that sulfation, and in particular, modification by chondroitin
sulfate is required for inducible hyaluronan binding in L cells. In the absence of fully processed
oligosaccharides, chondroitin sulfate is not essential for hyaluronan binding, indicating that the
effect of chondroitin sulfate is dependent on the glycosylation state of the cell. Thus, in addition to
glycosylation, chondroitin sulfate biosynthesis is an important post-translational modification that
can affect CD44-mediated hyaluronan binding.
CD44-hyaluronan interactions have been been implicated in leukocyte rolling and
extravasation at inflammatory sites. CD44 is normally expressed on leukocytes in an inactive state
that cannot bind hyaluronan but can be converted to an active state upon activation by antigen or
cytokines. Here it is demonstrated that the pro-inflammatory cytokine tumour necrosis factor α,
but not interferon γ, could convert CD44 from an inactive to an active hyaluronan binding form by
inducing the sulfation of CD44 on the SR91 myeloid cell line. This post-translational modification
was required for CD44-mediated binding to hyaluronan and to the vascular endothelial cell line,
SVEC4-10. Tumour necrosis factor α-inducible hyaluronan binding was also observed on
peripheral blood mononuclear cells, especially on CD14 positive monocytes and hyaluronan
binding appeared to be at least partially sulfation dependent. In addition, monocyte-specific pro-inflammatory
chemokines macrophage inflamatory protein-1α and monocyte chemoattractant
protein-1 could induce hyaluronan binding in a subpopulation of peripheral blood mononuclear
cells although it could not be determined definitively that the induction was sulfation dependent.
Taken together, sulfation may be a means of regulating CD44-mediated adhesion, both in
leukocytes at inflammatory sites as well as in fibroblasts to facilitate cell migration. === Science, Faculty of === Microbiology and Immunology, Department of === Graduate
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