PMID- 10395745
OWN - NLM
STAT- MEDLINE
DCOM- 19990812
LR  - 20061115
IS  - 0003-9861 (Print)
IS  - 0003-9861 (Linking)
VI  - 367
IP  - 2
DP  - 1999 Jul 15
TI  - Characterization of UDP-N-acetylglucosamine:alpha-6-d-mannoside
      beta-1,6-N-acetylglucosaminyltransferase V from a human hepatoma cell line Hep3B.
PG  - 281-8
AB  - UDP-N-acetylglucosamine:alpha-6-d-mannoside beta-1,
      6-N-acetylglucosaminyltransferase V (GlcNAcT-V) has been purified from cell
      extracts of the human hepatoma cell line, Hep3B, with 8.7% recovery. The purified
      enzymes had molecular masses of about 67 and 65 kDa on denaturated and natural
      conditions, respectively. The values of pI was 5.9. The GlcNAcT-V, when resolved 
      by SDS-PAGE, was positive for Schiff staining, suggesting that the enzyme is
      glycoprotein. When GlcN,GlcN-biant-PA and UDP-GlcNAc were used as substrates, the
      enzyme displayed a temperature optimum of around 50 degrees C and optimum an pH
      of 6.5. The enzyme was stable in response to incubation from pH 4.5 to pH 10.5 at
      4 degrees C for 24 h. The presence of UDP-GlcNAc and GlcN,GlcN-bi-PA protected
      the enzyme from heat inactivation, the extent depending upon the substrate
      concentration. The activity of the enzyme was stimulated by Mn2+ ion; however, it
      was inhibited by Fe3+. The enzyme activity was inhibited by another series of
      NDP-sugars including ADP-, CDP-, GDP-, and TDP-GlcNAc. Studies on the activity of
      the enzyme toward a variety of pyridylaminated sugars showed that the enzyme is
      most active toward biantennary (GlcN,GlcN-bi-PA) sugars. The enzymes had apparent
      Km values of 1.28 and 5.8 mM for GlcN,GlcN-bi-PA and UDP-GlcNAc, respectively. In
      order to isolate the GlcNAcT-V gene, PCR primers of GNN-1 and GNN-8 were designed
      and the amplified PCR product carrying the gene was cloned and sequenced.
      Nucleotide sequence analysis showed a 2220-bp open reading frame encoding a
      740-amino-acid protein. This was almost same as the previously reported human
      sequences, except for some sequence differences in three amino acids. The three
      amino acid changes were as follows: 375V --> L, 555T --> R, and 592A --> G. These
      studies represent the detailed characterization of a purified GlcNAcT-V from
      human hepatoma cell Hep3B.
CI  - Copyright 1999 Academic Press.
FAU - Park, C
AU  - Park C
AD  - College of Oriental Medicine, Dongguk University, Sukjang-Dong 707, Kyungju City,
      Kyungpook, 780-714, Korea.
FAU - Jin, U H
AU  - Jin UH
FAU - Lee, Y C
AU  - Lee YC
FAU - Cho, T J
AU  - Cho TJ
FAU - Kim, C H
AU  - Kim CH
LA  - eng
SI  - GENBANK/AF113921
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Arch Biochem Biophys
JT  - Archives of biochemistry and biophysics
JID - 0372430
RN  - 0 (Ions)
RN  - EC 2.4.1.- (N-Acetylglucosaminyltransferases)
RN  - EC 2.4.1.155 (alpha-1,6-mannosylglycoprotein beta
      1,6-N-acetylglucosaminyltransferase)
SB  - IM
MH  - Base Sequence
MH  - Carbohydrate Sequence
MH  - Carcinoma, Hepatocellular/*enzymology
MH  - Humans
MH  - Hydrogen-Ion Concentration
MH  - Ions
MH  - Kinetics
MH  - Models, Biological
MH  - Molecular Sequence Data
MH  - Mutation
MH  - N-Acetylglucosaminyltransferases/*chemistry/isolation & purification
MH  - Temperature
MH  - Tumor Cells, Cultured
EDAT- 1999/07/09 00:00
MHDA- 1999/07/09 00:01
CRDT- 1999/07/09 00:00
PHST- 1999/07/09 00:00 [pubmed]
PHST- 1999/07/09 00:01 [medline]
PHST- 1999/07/09 00:00 [entrez]
AID - 10.1006/abbi.1999.1252 [doi]
AID - S0003-9861(99)91252-3 [pii]
PST - ppublish
SO  - Arch Biochem Biophys. 1999 Jul 15;367(2):281-8. doi: 10.1006/abbi.1999.1252.