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Incorporation of N ‐acetylgalactosamine into consecutive threonine residues in MUC2 tandem repeat by recombinant human N ‐acetyl‐ D ‐galactosamine transferase‐T1, T2 and T3
Author(s) -
Iida Shin-ichiro,
Takeuchi Hideyuki,
Hassan Helle,
Clausen Henrik,
Irimura Tatsuro
Publication year - 1999
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(99)00445-7
Subject(s) - peptide , chemistry , galactosamine , biochemistry , oligopeptide , enzyme , tandem mass spectrometry , chromatography , mass spectrometry , glucosamine
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.