Localization of human heparan glucosaminyl N-deacetylase/N-sulphotransferase to the trans-Golgi network.
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.
Humphries
D E
DE
Department of Veterans Affairs Medical Center, Department of Medicine, Harvard Medical School, Boston, MA 02130, USA.
Sullivan
B M
BM
Aleixo
M D
MD
Stow
J L
JL
D000581:Amidohydrolases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D015153:Blotting, Western; D020126:Brefeldin A; D002460:Cell Line; D003001:Cloning, Molecular; D003517:Cyclopentanes / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000166:cytology / Q000201:enzymology; D000939:Epitopes / Q000737:chemistry / Q000276:immunology; D005455:Fluorescent Antibody Technique; D006056:Golgi Apparatus / Q000201:enzymology*; D006493:Heparin / Q000096:biosynthesis; D006497:Heparan Sulfate / Q000096:biosynthesis; D006801:Humans; D051379:Mice; D008856:Microscopy, Fluorescence; D009842:Oligopeptides; D010455:Peptides / Q000276:immunology / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis; D015238:Sulfotransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014471:Umbilical Veins
D000581:Amidohydrolases; D000911:Antibodies, Monoclonal; D020126:Brefeldin A; D003517:Cyclopentanes; D000939:Epitopes; D006493:Heparin; D006497:Heparan Sulfate; D009842:Oligopeptides; D010455:Peptides; D012333:RNA, Messenger; D015238:Sulfotransferases; C085706:FLAG peptide; C021350:heparitin sulfotransferase