Cloning of a human UDP-N-acetyl-α-D-galactosamine:polypeptide N-acetylgalactosaminyltransferase that complements other GalNAc-transferases in complete O-glycosylation of the MUC1 tandem repeat

被引:187
作者
Bennett, EP
Hassan, H
Mandel, U
Mirgorodskaya, E
Roepstorff, P
Burchell, J
Taylor-Papadimitriou, J
Hollingsworth, MA
Merkx, G
van Kessel, AG
Eiberg, H
Steffensen, R
Clausen, H
机构
[1] Univ Copenhagen, Sch Dent, Fac Hlth Sci, DK-2200 Copenhagen N, Denmark
[2] Odense Univ, Dept Mol Biol, DK-5230 Odense, Denmark
[3] Imperial Canc Res Fund, London WC2A 3PX, England
[4] Univ Nebraska, Med Ctr, Eppley Inst Res Canc & Allied Dis, Omaha, NE 68198 USA
[5] Univ Nijmegen Hosp, Dept Human Genet, NL-6500 HB Nijmegen, Netherlands
[6] Univ Copenhagen, Fac Hlth Sci, Genet Inst, DK-2200 Copenhagen N, Denmark
[7] Aalborg Hosp, Reg Ctr Blood Transfus, Aalborg, Denmark
关键词
D O I
10.1074/jbc.273.46.30472
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
A fourth human UDP-GalNAc:polypeptide N- acetylgalactosaminyltransferase, designated GalNAc-T4, was cloned and expressed. The genomic organization of GalNAc-T4 is distinct from GalNAc-T1, -T2, and -T3, which contain multiple coding exons, in that the coding region is contained in a single exon. GalNAc-T4 was placed at human chromosome 12q21.3-q22 by in situ hybridization and linkage analysis. GalNAc-T4 expressed in Sf9 cells or in a stably transfected Chinese hamster ovary cell line exhibited a unique acceptor substrate specificity. GalNAc-T4 transferred GalNAc to two sites in the MUC1 tandem repeat sequence (Ser in GVTSA and Thr in PDTR) using a 24-mer glycopeptide with GalNAc residues attached at sites utilized by GalNAc-T1, -T2, and -T3 (TAPPAHGVTSAPDTRPAPGSTAPPA, GalNAc attachment sites underlined). Furthermore, GalNAc-T4 showed the best kinetic properties with an O-glycosylation site in the P-selectin glycoprotein ligand-1 molecule. Northern analysis of human organs revealed a wide expression pattern. Immunohistology with a monoclonal antibody showed the expected Golgi-like localization in salivary glands. A single base polymorphism, G1516A (Val to lie), was identified (allele frequency 34%). The function of GalNAc-T4 complements other GalNAc-transferases in O-glycosylation of MUC1 showing that glycosylation of MUC1 is a highly ordered process and changes in the repertoire or topology of GalNAc-transferases will result in altered pattern of O-glycan attachments.
引用
收藏
页码:30472 / 30481
页数:10
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