Isolation, characterization, and expression of a cDNA encoding N- acetylglucosaminyltransferase V

M. Shoreibah, G. S. Perng, B. Adler, J. Weinstein, R. Basu, R. Cupples, D. Wen, J. K. Browne, P. Buckhaults, N. Fregien, M. Pierce

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111 Scopus citations

Abstract

A cDNA clone for the complete coding sequence for α-1,3(6)-mannosylglycoprotein β-1,6-N-acetylglucosaminyltransferase V (GlcNAc-T V, EC 2.4.1.155) was isolated and expressed in COS-7 cells. Degenerate oligonucleotide primers for polymerase chain reaction were synthesized based on the amino acid sequence of three tryptic peptides isolated from affinity-purified GlcNAc-T V. Polymerase chain reaction amplimers were isolated from rat and mouse mRNA. A cDNA-encoding full-length enzyme was isolated from a rat 1 cell (EJ-ras-transformed) library and sequenced. Transient expression of this clone in COS-7 cells, followed by enzymatic activity assays, demonstrated that this cDNA sequence encodes GlcNAc-T V. Northern analysis of rat kidney mRNA revealed a single band corresponding to a length of about 7 kilobases. Sequence analysis of the cDNA clone demonstrated an open reading frame that encoded a type II membrane protein of 740 amino acids.

Original languageEnglish (US)
Pages (from-to)15381-15385
Number of pages5
JournalJournal of Biological Chemistry
Volume268
Issue number21
StatePublished - Jan 1 1993

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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    Shoreibah, M., Perng, G. S., Adler, B., Weinstein, J., Basu, R., Cupples, R., Wen, D., Browne, J. K., Buckhaults, P., Fregien, N., & Pierce, M. (1993). Isolation, characterization, and expression of a cDNA encoding N- acetylglucosaminyltransferase V. Journal of Biological Chemistry, 268(21), 15381-15385.