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

Mohamed Shoreibah, Guang Shing Perng, Beverly Adler, Jasminder Weinstein, Rita Basu, Rod Cupples, Duanzhi Wen, Jeffrey K. Browne, Phillip Buckhaults, Nevis L. Fregien, Michael Pierce

Research output: Contribution to journalArticle

110 Citations (Scopus)

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
Pages (from-to)15381-15385
Number of pages5
JournalJournal of Biological Chemistry
Volume268
Issue number21
StatePublished - Jul 25 1993

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alpha-1,6-mannosylglycoprotein beta 1,6-N-acetylglucosaminyltransferase
Complementary DNA
Rats
Clone Cells
COS Cells
Polymerase chain reaction
Amino Acids
Polymerase Chain Reaction
Messenger RNA
DNA Primers
Enzyme Assays
Open Reading Frames
Libraries
Sequence Analysis
Amino Acid Sequence
Assays
Membrane Proteins
Kidney
Peptides
Enzymes

ASJC Scopus subject areas

  • Biochemistry

Cite this

Shoreibah, M., Perng, G. S., Adler, B., Weinstein, J., Basu, R., Cupples, R., ... Pierce, M. (1993). Isolation, characterization, and expression of a cDNA encoding n-acetylglucosaminyltransferase V. Journal of Biological Chemistry, 268(21), 15381-15385.

Isolation, characterization, and expression of a cDNA encoding n-acetylglucosaminyltransferase V. / Shoreibah, Mohamed; Perng, Guang Shing; Adler, Beverly; Weinstein, Jasminder; Basu, Rita; Cupples, Rod; Wen, Duanzhi; Browne, Jeffrey K.; Buckhaults, Phillip; Fregien, Nevis L.; Pierce, Michael.

In: Journal of Biological Chemistry, Vol. 268, No. 21, 25.07.1993, p. 15381-15385.

Research output: Contribution to journalArticle

Shoreibah, M, Perng, GS, Adler, B, Weinstein, J, Basu, R, Cupples, R, Wen, D, Browne, JK, Buckhaults, P, Fregien, NL & Pierce, M 1993, 'Isolation, characterization, and expression of a cDNA encoding n-acetylglucosaminyltransferase V', Journal of Biological Chemistry, vol. 268, no. 21, pp. 15381-15385.
Shoreibah M, Perng GS, Adler B, Weinstein J, Basu R, Cupples R et al. Isolation, characterization, and expression of a cDNA encoding n-acetylglucosaminyltransferase V. Journal of Biological Chemistry. 1993 Jul 25;268(21):15381-15385.
Shoreibah, Mohamed ; Perng, Guang Shing ; Adler, Beverly ; Weinstein, Jasminder ; Basu, Rita ; Cupples, Rod ; Wen, Duanzhi ; Browne, Jeffrey K. ; Buckhaults, Phillip ; Fregien, Nevis L. ; Pierce, Michael. / Isolation, characterization, and expression of a cDNA encoding n-acetylglucosaminyltransferase V. In: Journal of Biological Chemistry. 1993 ; Vol. 268, No. 21. pp. 15381-15385.
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