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J Biol Chem, Vol. 274, Issue 8, 4504-4512, February 19, 1999
1,6-N-ACETYLGLUCOSAMINYLTRANSFERASE FORMING CORE 2 AND
CORE 4
,
,,,
From the A novel human
UDP-GlcNAc:Gal/GlcNAc School of Dentistry, University of
Copenhagen, Nørre Allé 20, 2200 Copenhagen N, Denmark, the
§ Eppley Institute for Research in Cancer and Allied
Diseases, University of Nebraska Medical Center, Omaha, Nebraska 68198, the ¶ University of Georgia, Complex Carbohydrate Research Center,
220 Riverbend Road, Athens, Georgia 30602, and the Department of
Human Genetics, University Hospital Nijmegen,
6500 HB Nijmegen, The Netherlands
1-3GalNAc
1,6GlcNAc-transferase,
designated C2/4GnT, was identified by BLAST analysis of expressed
sequence tags. The sequence of C2/4GnT encoded a putative type II
transmembrane protein with significant sequence similarity to human
C2GnT and IGnT. Expression of the secreted form of C2/4GnT in insect
cells showed that the gene product had UDP-N-acetyl--D-glucosamine:acceptor
1,6-N-acetylglucosaminyltransferase (1,6GlcNAc-transferase) activity. Analysis of substrate specificity revealed that the enzyme catalyzed O-glycan branch
formation of the core 2 and core 4 type. NMR analyses of the product
formed with core 3-para-nitrophenyl confirmed the product
core 4-para-nitrophenyl. The coding region of C2/4GnT was
contained in a single exon and located to chromosome 15q21.3. Northern
analysis revealed a restricted expression pattern of C2/4GnT mainly in
colon, kidney, pancreas, and small intestine. No expression of C2/4GnT
was detected in brain, heart, liver, ovary, placenta, spleen, thymus,
and peripheral blood leukocytes. The expression of core 2 O-glycans has been correlated with cell differentiation
processes and cancer. The results confirm the predicted existence of a
1,6GlcNAc-transferase that functions in both core 2 and core 4 O-glycan branch formation. The redundancy in
1,6GlcNAc-transferases capable of forming core 2 O-glycans is important for understanding the mechanisms
leading to specific changes in core 2 branching during cell development and malignant transformation.
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