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PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
9
pubmed:dateCreated
1998-3-25
pubmed:databankReference
pubmed:abstractText
The HNK-1 carbohydrate is expressed on various adhesion molecules in the nervous system and is suggested to play a role in cell-cell and cell-substratum interactions. Here we describe the isolation and functional expression of a cDNA encoding a human sulfotransferase that synthesizes the HNK-1 carbohydrate epitope. A mutant Chinese hamster ovary cell line, Lec2, which stably expresses human neural cell adhesion molecule (N-CAM) (Lec2-NCAM), was first established. Lec2-NCAM was co-transfected with a human fetal brain cDNA library, a cDNA encoding the rat glucuronyltransferase that forms a precursor of the HNK-1 carbohydrate, and a vector encoding the polyoma large T antigen. The transfected Lec2-NCAM cells expressing the HNK-1 glycan were enriched by fluorescence-activated cell sorting. Sibling selection of recovered plasmids resulted in a cDNA encoding a sulfotransferase, HNK-1ST, that directs the expression of the HNK-1 carbohydrate epitope on the cell surface. The deduced amino acid sequence indicates that the enzyme is a type II membrane protein. Sequence analysis revealed that there is a short amino acid sequence in the presumed catalytic domain, which is highly homologous to the corresponding sequence in other Golgi-associated sulfotransferases so far cloned. The amount of HNK-1ST transcript is high in fetal brain compared with fetal lung, kidney, and liver. Expression of HNK-1ST resulted in the formation of the HNK-1 epitope on N-CAM and a soluble chimeric form of HNK-1ST was shown to add a sulfate group to a precursor, GlcAbeta1-->3Galbeta1-->4GlcNAcbeta1-->R, forming sulfo-->3GlcAbeta1-->3Galbeta1-->4GlcNAcbeta1-->R. The results combined together indicate that the cloned HNK-1ST directs the synthesis of the HNK-1 carbohydrate epitope on both glycoproteins and glycolipids in the nervous tissues.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Feb
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
27
pubmed:volume
273
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
5190-5
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed-meshheading:9478973-Amino Acid Sequence, pubmed-meshheading:9478973-Animals, pubmed-meshheading:9478973-Base Sequence, pubmed-meshheading:9478973-Brain, pubmed-meshheading:9478973-CHO Cells, pubmed-meshheading:9478973-Carbohydrate Sequence, pubmed-meshheading:9478973-Cell Adhesion, pubmed-meshheading:9478973-Cloning, Molecular, pubmed-meshheading:9478973-Cricetinae, pubmed-meshheading:9478973-DNA, Complementary, pubmed-meshheading:9478973-Epitopes, pubmed-meshheading:9478973-Fetus, pubmed-meshheading:9478973-Gene Expression, pubmed-meshheading:9478973-Glycolipids, pubmed-meshheading:9478973-Golgi Apparatus, pubmed-meshheading:9478973-Humans, pubmed-meshheading:9478973-Molecular Sequence Data, pubmed-meshheading:9478973-Neural Cell Adhesion Molecules, pubmed-meshheading:9478973-Rats, pubmed-meshheading:9478973-Recombinant Proteins, pubmed-meshheading:9478973-Sequence Analysis, DNA, pubmed-meshheading:9478973-Sequence Homology, Amino Acid, pubmed-meshheading:9478973-Sulfotransferases, pubmed-meshheading:9478973-Tissue Distribution, pubmed-meshheading:9478973-Trisaccharides
pubmed:year
1998
pubmed:articleTitle
Expression cloning of a human sulfotransferase that directs the synthesis of the HNK-1 glycan on the neural cell adhesion molecule and glycolipids.
pubmed:affiliation
Glycobiology Program, La Jolla Cancer Research Center, The Burnham Institute, La Jolla, California 92037, USA.
pubmed:publicationType
Journal Article, Comparative Study, Research Support, U.S. Gov't, P.H.S., Research Support, Non-U.S. Gov't