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J Biol Chem, Vol. 273, Issue 44, 28878-28888, October 30, 1998
From the Abundant hyaluronan is present between epidermal
keratinocytes. However, virtually nothing is known regarding its
organization in the limited extracellular space between these cells. We
have used metabolic labeling with [3H]glucosamine
and [35S]sulfate and a hyaluronan-specific biotinylated
probe to study the metabolism of hyaluronan and its localization in
monolayer cultures of a rat epidermal keratinocyte cell line.
Hyaluronan (~20 fg/cell) was present on the apical and lateral
surfaces of the cells in two nearly equal pools, either in patches
(~160/cell) or diffusely spread. The hyaluronan in the patches is
bound to CD44 as indicated by co-localization with an antibody to CD44, and by displacement with hyaluronan decasaccharides as well as with an
antibody that blocks hyaluronan binding to CD44. The inability of
hyaluronan oligomers shorter than 10 monosaccharides to displace hyaluronan suggests that CD44 dimerization or cooperative interactions are required for tight binding. The diffuse hyaluronan pool is likely
bound to hyaluronan synthase during its biosynthesis.
Hyaluronan Bound to CD44 on Keratinocytes Is Displaced by
Hyaluronan Decasaccharides and Not Hexasaccharides
§,
,,, and§
Department of Anatomy, University of Kuopio,
FIN-70211 Kuopio, Finland, the § Department of Biomedical
Engineering, Connective Tissue Biology Section, Cleveland Clinic
Research Institute, Cleveland, Ohio 44195, and the Department
of Anatomy and Cell Biology, University of Michigan,
Ann Arbor, Michigan 48109
Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.
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