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PMID: 9230113 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Localization of human heparan glucosaminyl N-deacetylase/N-sulphotransferase to the trans-Golgi network.

The Biochemical journal ·Vol. 325 ( Pt 2) ·1997-07-15 ·Pages 351-7

Humphries DE, Sullivan BM, Aleixo MD, Stow JL

Abstract

In order to determine the intracellular location of heparan N-deacetylase/N-sulphotransferase, cDNAs encoding human heparan glucosaminyl N-deacetylase/N-sulphotransferase were cloned from human umbilical vein endothelial cells. The deduced amino acid sequence was identical to that of the human heparan N-sulphotransferase cloned previously [Dixon, Loftus, Gladwin, Scambler, Wasmuth and Dixon (1995) Genomics 26, 239-244]. RNA blot analysis indicated that two heparan N-sulphotransferase transcripts of approx. 8.5 and 4 kb were produced in all tissues. Expression was most abundant in heart, liver and pancreas. A cDNA encoding a Flag-tagged human heparan N-sulphotransferase (where Flag is an epitope with the sequence DYKDDDDK) was transfected into mouse LTA cells. Immunofluorescence detection using anti-Flag monoclonal antibodies demonstrated that the enzyme was localized to the trans-Golgi network. A truncated Flag-tagged heparan N-sulphotransferase was also retained in the Golgi, indicating that, as for many other Golgi enzymes, the N-terminal region of heparan N-sulphotransferase is sufficient for retention in the Golgi apparatus.

MeSH Terms
Amidohydrolases/chemistry,genetics,metabolism Animals Antibodies, Monoclonal/immunology Blotting, Western Brefeldin A Cell Line Cloning, Molecular Cyclopentanes/pharmacology Endothelium, Vascular/cytology,enzymology Epitopes/chemistry,immunology Fluorescent Antibody Technique Golgi Apparatus/enzymology Heparin/biosynthesis Heparitin Sulfate/biosynthesis Humans Mice Microscopy, Fluorescence Oligopeptides Peptides/immunology,metabolism RNA, Messenger/analysis Sulfotransferases/chemistry,genetics,metabolism Umbilical Veins
Chemicals
Antibodies, Monoclonal Cyclopentanes Epitopes Oligopeptides Peptides RNA, Messenger Brefeldin A Heparin Heparitin Sulfate FLAG peptide Sulfotransferases heparitin sulfotransferase Amidohydrolases
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Humphries D E
Department of Veterans Affairs Medical Center, Department of Medicine, Harvard Medical School, Boston, MA 02130, USA.
Sullivan B M
Aleixo M D
Stow J L
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj3250351
Published
1997-07-15
Pages
351-7
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1218567
Subset
IM
Grants
NIDDK NIH HHS · DK42881 · United States
Databases
GENBANK
U36600
External Links
PubMed source DOI: 10.1042/bj3250351 PMC Full Text View journal details
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