title: Isolation, characterization, and expression of a cDNA encoding N-acetylglucosaminyltransferase V.
abstract: A cDNA clone for the complete coding sequence for alpha-1,3(6)-mannosylglycoprotein beta-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.
authors.affiliation: Department of Biochemistry, University of Georgia, Athens 30602.
authors.forename: M
authors.initials: M
authors.lastname: Shoreibah
authors.forename: G S
authors.initials: GS
authors.lastname: Perng
authors.forename: B
authors.initials: B
authors.lastname: Adler
authors.forename: J
authors.initials: J
authors.lastname: Weinstein
authors.forename: R
authors.initials: R
authors.lastname: Basu
authors.forename: R
authors.initials: R
authors.lastname: Cupples
authors.forename: D
authors.initials: D
authors.lastname: Wen
authors.forename: J K
authors.initials: JK
authors.lastname: Browne
authors.forename: P
authors.initials: P
authors.lastname: Buckhaults
authors.forename: N
authors.initials: N
authors.lastname: Fregien
authors.forename: M
authors.initials: M
authors.lastname: Pierce
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D004247:DNA / Q000302:isolation & purification; D007668:Kidney / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D017351:N-Acetylglucosaminyltransferases / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats
chemical_list: D012333:RNA, Messenger; D004247:DNA; D017351:N-Acetylglucosaminyltransferases; C052254:alpha-1,6-mannosylglycoprotein beta 1,6-N-acetylglucosaminyltransferase