PMID- 10430883
OWN - NLM
STAT- MEDLINE
DCOM- 19990909
LR  - 20190501
IS  - 0027-8424 (Print)
IS  - 0027-8424 (Linking)
VI  - 96
IP  - 16
DP  - 1999 Aug 3
TI  - Expression cloning of a human alpha1, 4-N-acetylglucosaminyltransferase that
      forms GlcNAcalpha1-->4Galbeta-->R, a glycan specifically expressed in the gastric
      gland mucous cell-type mucin.
PG  - 8991-6
AB  - Among mucus-secreting cells, the gastric gland mucous cells, Brunner's glands,
      accessory glands of pancreaticobiliary tract, and pancreatic ducts exhibiting
      gastric metaplasia are unique in that they express class III mucin identified by 
      paradoxical Con A staining composed of periodate oxidation, sodium borohydride
      reduction, Con A, and horseradish peroxidase reaction. Recently it was shown that
      these mucous cells secrete glycoproteins having GlcNAcalpha1-->4Galbeta-->R at
      nonreducing terminals of the carbohydrate moieties. Herein we describe the
      expression cloning of a cDNA encoding a human
      alpha1,4-N-acetylglucosaminyltransferase (alpha4GnT), a key enzyme for the
      formation of GlcNAcalpha1-->4Galbeta1-->R. COS-1 cells were thus cotransfected
      with a stomach cDNA library and a leukosialin cDNA. Transfected COS-1 cells were 
      screened by using monoclonal antibodies specific for GlcNAcalpha1-->4Galbeta-->R 
      and enriched by fluorescence-activated cell sorting. Sibling selection of
      recovered plasmids resulted in a cDNA clone that directs the expression of
      GlcNAcalpha1-->4Galbeta-->R. The deduced amino acid sequence predicts a type II
      membrane protein with 340 amino acids, showing no significant similarity with any
      other proteins. The alpha4GnT gene is located at chromosome 3p14.3, and its
      transcripts are expressed in the stomach and pancreas. An in vitro GlcNAc
      transferase assay by using a soluble alpha4GnT revealed that alpha1,4-linked
      GlcNAc residues are transferred most efficiently to core 2 branched O-glycans
      (Galbeta1-->4GlcNAcbeta1-->6(Galbeta1-->3)GalNAc), forming
      GlcNAcalpha1-->4Galbeta1-->4GlcNAcbeta1-->6(GlcNAca lpha1-->4Galbeta1-
      ->3)GalNAc. Transfection of alpha4GnT cDNA into gastric adenocarcinoma AGS cells 
      produced class III mucin, indicating that alpha4GnT is responsible for the
      formation of class III Con A reactivity. These results indicate that the
      alpha4GnT is a glycosyltransferase that forms alpha1,4-linked GlcNAc residues,
      preferentially in O-glycans.
FAU - Nakayama, J
AU  - Nakayama J
AD  - Department of Laboratory Medicine, Shinshu University School of Medicine,
      Matsumoto 390-8621, Japan. jun@hsp.md.shinshu-u.ac.jp
FAU - Yeh, J C
AU  - Yeh JC
FAU - Misra, A K
AU  - Misra AK
FAU - Ito, S
AU  - Ito S
FAU - Katsuyama, T
AU  - Katsuyama T
FAU - Fukuda, M
AU  - Fukuda M
LA  - eng
SI  - GENBANK/AF141315
GR  - CA48737/CA/NCI NIH HHS/United States
GR  - P01 CA071932/CA/NCI NIH HHS/United States
GR  - R01 CA048737/CA/NCI NIH HHS/United States
GR  - R01 CA033000/CA/NCI NIH HHS/United States
GR  - CA33000/CA/NCI NIH HHS/United States
GR  - CA71932/CA/NCI NIH HHS/United States
GR  - R37 CA033000/CA/NCI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Proc Natl Acad Sci U S A
JT  - Proceedings of the National Academy of Sciences of the United States of America
JID - 7505876
RN  - 0 (Polysaccharides)
RN  - 0 (Recombinant Proteins)
RN  - EC 2.4.1.- (N-Acetylglucosaminyltransferases)
SB  - IM
MH  - Adult
MH  - Amino Acid Sequence
MH  - Animals
MH  - Base Sequence
MH  - COS Cells
MH  - Carbohydrate Conformation
MH  - Carbohydrate Sequence
MH  - Cell Membrane/enzymology
MH  - Chromosome Mapping
MH  - *Chromosomes, Human, Pair 3
MH  - Cloning, Molecular
MH  - Female
MH  - Gastric Mucosa/cytology/*enzymology
MH  - Gene Expression Regulation, Enzymologic
MH  - Humans
MH  - Male
MH  - Molecular Sequence Data
MH  - N-Acetylglucosaminyltransferases/biosynthesis/chemistry/*genetics
MH  - Organ Specificity
MH  - Polysaccharides/*biosynthesis/chemistry
MH  - Recombinant Proteins/biosynthesis/chemistry
MH  - Substrate Specificity
MH  - Transfection
PMC - PMC17720
EDAT- 1999/08/04 00:00
MHDA- 1999/08/04 00:01
CRDT- 1999/08/04 00:00
PHST- 1999/08/04 00:00 [pubmed]
PHST- 1999/08/04 00:01 [medline]
PHST- 1999/08/04 00:00 [entrez]
AID - 10.1073/pnas.96.16.8991 [doi]
PST - ppublish
SO  - Proc Natl Acad Sci U S A. 1999 Aug 3;96(16):8991-6. doi: 10.1073/pnas.96.16.8991.