Core O-glycans required for lymphocyte homing gene knockout mice of core 1 beta1,3-N-acetylglucosaminyltransferase and core 2 N-acetylglucosaminyltransferase.
Mucin-type O-glycans are synthesized by sequential reaction of glycosyltransferases that have different substrate specificities. To know the significance of specific O-glycan structures, many researchers have been making mice deficient in corresponding enzymes for the synthesis of the O-glycan structures. Here we describe the analysis of gene knockout mice of core 2 branching enzyme (core 2 N-acetylglucosaminyltransferase, Core2GlcNAcT) and core 1 extension enzyme (core 1 beta1,3-N-acetylglucosaminyltransferase, Core1-beta3GlcNAcT). Because mucin-type O-glycans present sialyl Lewis X (sLeX) and sulfated version of the glycans, which are L-selectin ligands, at the reducing end, the amounts of the ligands of these knockout mice would be reduced. The methods described here are to analyze the interaction between L-selectin and its ligand 6-sulfo sLeX such as lymphocyte homing assay, staining of frozen section, and blotting using L- and E-selectin-IgM chimeric proteins.
Mitoma
Junya
J
Division of Glyco-Signal Research, Institute of Molecular Biomembrane and Glycobiology, Tohoku Pharmaceutical University, Komatsushima, Aoba, Sendai, Japan.
Fukuda
Minoru
M
D000818:Animals; D018390:Gene Targeting; D019041:L-Selectin / Q000378:metabolism*; D008024:Ligands; D051379:Mice; D018345:Mice, Knockout; D017351:N-Acetylglucosaminyltransferases / Q000235:genetics* / Q000378:metabolism*; D011134:Polysaccharides / Q000378:metabolism*; D016201:Receptors, Lymphocyte Homing / Q000235:genetics* / Q000378:metabolism
D008024:Ligands; D011134:Polysaccharides; D016201:Receptors, Lymphocyte Homing; D019041:L-Selectin; D017351:N-Acetylglucosaminyltransferases