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Blood, 15 June 2001, Vol. 97, No. 12, pp. 3806-3811
HEMOSTASIS, THROMBOSIS, AND VASCULAR BIOLOGY
Differential requirements for the O-linked branching enzyme core
2  1-6-N-glucosaminyltransferase in biosynthesis of ligands for
E-selectin and P-selectin
Karen R. Snapp,
Christine
E. Heitzig,
Lesley G. Ellies,
Jamey D. Marth, and
Geoffrey S. Kansas
From the Department of Microbiology-Immunology,
Northwestern University Medical School, Chicago, IL, and the Howard
Hughes Medical Institute and Department of Cellular and Molecular
Medicine, University of California San Diego, La Jolla, CA.
Selectins are carbohydrate-binding adhesion molecules that play
important roles in control of leukocyte traffic. Glycosyltransferases involved in selectin ligand biosynthesis include the
 1,3-fucosyltransferases FucT-VII and FucT-IV, one or more
sialyltransferases, and at least one O-linked branching enzyme.
Previous studies have shown that core 2  1-6-N-glucosaminyltransferase (C2GlcNAcT-I; EC 2.4.1.102) is required for functional modification of PSGL-1, the leukocyte P-selectin ligand, but have been ambiguous on whether this enzyme is
involved in E-selectin ligand formation. Using an attachment and
rolling assay under defined shear flow in vitro, this study shows that
C2GlcNAcT-I lymphoid cells stably transfected with
FucT-VII complementary DNA attach and roll well on E-selectin at 1.5 dynes/cm.2 Further, attachment and rolling on P-selectin
of neutrophils is sharply reduced and that of short- term
polarized Th1 cells is virtually abolished, with leukocytes from
C2GlcNAcT-I/ mice. In contrast, both
neutrophils and Th1 cells from C2GlcNAcT-I/ mice
attach and roll as well as wild-type cells on E-selectin. These results
show that C2GlcNAcT-I is selectively required for biosynthesis of
ligands for P-selectin, but is not essential for at least some
E-selectin ligands. Distinct requirements for C2GlcNAcT-I in the
formation of ligands for E-selectin versus P-selectin represents a
novel level of regulation of expression of selectin ligands and
lymphocyte traffic.
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