Incorporation of N‐acetylgalactosamine into consecutive threonine residues in MUC2 tandem repeat by recombinant human N‐acetyl‐D‐galactosamine transferase‐T1, T2 and T3

FEBS Letters

Takeuchi, Hideyuki²  Irimura, Tatsuro²  Hassan, Helle¹  Clausen, Henrik¹  Iida, Shin-ichiro² 

[1] Department of Oral Diagnostics, Faculty of Health Sciences, School of Dentistry, University of Copenhagen, Nørre Alle 20, DK-2200 Copenhagen, Denmark;Laboratory of Cancer Biology and Molecular Immunology, Graduate school of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan

关键词: Mucin;    N-acetylgalactosaminyltransferase;    O-glycosylation;    MUC2;    Tn antigen;    GalNAc;    N-acetyl-D-galactosamine;    GalNAc-T;    UDP-GalNAc:polypeptide α-GalNAc-transferase (EC 2.4.1.41);    HPLC;    high performance liquid chromatography;    MALDI-TOF MS;    matrix-assisted laser desorption ionization time of flight mass spectrometry;    PTH;    phenylthiohydantoin;    TFA;    trifluoroacetic acid;   

DOI  :  10.1016/S0014-5793(99)00445-7

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

An oligopeptide containing three consecutive Thr residues mimicking the tandem repeat portion of MUC2 (PTTTPLK) was investigated for the acceptor specificity to UDP-N-acetyl-D-galactosamine:peptide N-acetylgalactosaminyltransferase isozymes, UDP-N-acetyl-D-galactosamine:peptide N-acetylgalactosaminyltransferase-T1, T2 and T3. The enzymatic reaction products were fractionated by the reversed-phase high performance liquid chromatography, then characterized by matrix-assisted laser desorption ionization time of flight mass spectrometry and by a peptide sequencing analysis. A maximum of two, one or three N-acetyl-D-galactosamine residues was transferred by UDP-N-acetyl-D-galactosamine:peptide N-acetylgalactosaminyltransferase-T1, T2 or T3, respectively. The preferential orders of N-acetyl-D-galactosamine incorporation were Thr-2, then Thr-4 for UDP-N-acetyl-D-galactosamine:peptide N-acetylgalactosaminyltransferase-T1, Thr-2 for UDP-N-acetyl-D-galactosamine:peptide N-acetylgalactosaminyltransferase-T2 and Thr-4, Thr-3, then Thr-2 for UDP-N-acetyl-D-galactosamine:peptide N-acetylgalactosaminyltransferase-T3.

【 授权许可】

Unknown