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J. Biol. Chem., Vol. 278, Issue 13, 11150-11158, March 28, 2003
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From the CD44 can function as an adhesion receptor that
mediates leukocyte rolling on hyaluronan (HA). To study the
contributions of different domains of the standard isoform of CD44 to
cell rolling, a CD44-negative mouse T lymphoma AKR1 was transfected
with wild type (WT) or mutated cDNA constructs. A parallel flow
chamber was used to study the rolling behavior of CD44 transfectants on immobilized HA. For CD44WT transfectants, the fraction of cells that
rolled and the rolling velocity was inversely proportional to the
amount of cell surface CD44. When the cytoplasmic domain distal to
Gly305 or sequences that serve as binding sites for
cytoskeletal linker proteins, were deleted or replaced with foreign
sequences, no significant changes in the rolling behavior of mutant
cells, compared with the transfectant expressing CD44WT, were observed.
Transfectants lacking 64 amino acids of the cytoplasmic tail distal to
Cys295 adhered to HA but showed enhanced rolling at low
shear forces. When 83 amino acids from the "non-conserved"
membrane-proximal region of the CD44 extracellular domain were deleted,
cells adhered firmly to the HA substrate and did not roll at any fluid
shear force tested. Unlike wild type cells that exhibited a nearly
homogenous distribution of CD44 on a smooth cell surface, cells
expressing the non-conserved region deletion mutant accumulated CD44 in
membrane protrusions. Disruption of the actin cytoskeleton with
cytochalasin B precluded the formation of membrane protrusions,
however, treated cells still adhered firmly to HA and did not roll. We
conclude that interaction between the cytoplasmic domain of CD44 and
the cytoskeleton is not required for cell rolling on immobilized
ligand. The strong effect of deletion of the non-conserved region of
the extracellular domain argues for a critical role of this region in
CD44-dependent rolling and adhesion interactions with HA
under flow.
Role of the Extracellular and Cytoplasmic Domains of CD44 in the
Rolling Interaction of Lymphoid Cells with Hyaluronan under Physiologic
Flow*
,,,,¶
Department of Orthopedic Surgery, Section of
Biochemistry and Molecular Biology, Rush University at
Rush-Presbyterian-St. Luke's Medical Center, Chicago, Illinois 60612 and the § Molecular and Cell Biology Laboratory, The Salk
Institute, San Diego, California 92186
*
This work was supported in part by Grants AI-31613 (to
R. H.) and AR-45652 (to K. M.) from the National Institutes of
Health.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
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