Molecular cloning and expression of cDNA encoding chicken UDP-N-acetyl-D-glucosamine (GlcNAc): GlcNAcβ1-6(GlcNAcβ1-2)-Manα1-R[GlcNAc to Man]β1,4N-acetylglucosaminyltransferase VI

Y. Sakamoto, T. Taguchi, K. Honke, H. Korekane, H. Watanabe, Y. Tano, N. Dohmae, K. Takio, A. Horii, N. Taniguchi

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

A cDNA that encodes UDP-N-acetyl-D-glucosamine (GlcNAc):GlcNAcβ1-6(GlcNAcβ1-2)Manα1-R[GlcNAc to Man]β1,4N-acetylglucosaminyltransferase VI (GnT VI), which is responsible for the formation of pentaantennary asparagine-linked oligosaccharides (N-glycans), has been cloned from a hen oviduct cDNA library based on the partial amino acid sequences of the purified enzyme. The isolated cDNA clone contained an open reading frame encoding 464 amino acids, including all of the peptides that were sequenced. The deduced amino acid sequence predicts a type II transmembrane topology and contains two potential N-glycosylation sites. The primary structure was found to be significantly similar to human GnT IV-homologue, the gene for which was cloned from the deleted region in pancreatic cancer, and to human and bovine GnT IVs. Chicken GnT VI-transfected COS-1 cells showed a high GnT VI activity (26.8 pmol/h/mg protein), whereas nontransfected, mocktransfected, or human GnT IV-homologue-transfected COS-1 cells had no activity. Northern blot analysis using poly(A)+ RNA from hen oviduct indicated that the size of GnT VI mRNA is 2.1 kilobases. Reverse transcription-polymerase chain reaction analysis showed that GnT VI mRNA was relatively highly expressed in oviduct, spleen, lung, and colon.

Original languageEnglish
Pages (from-to)36029-36034
Number of pages6
JournalJournal of Biological Chemistry
Volume275
Issue number46
DOIs
Publication statusPublished - 2000 Nov 17

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Fingerprint Dive into the research topics of 'Molecular cloning and expression of cDNA encoding chicken UDP-N-acetyl-D-glucosamine (GlcNAc): GlcNAcβ1-6(GlcNAcβ1-2)-Manα1-R[GlcNAc to Man]β1,4N-acetylglucosaminyltransferase VI'. Together they form a unique fingerprint.

  • Cite this