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Purification and characterization of 3'-phosphoadenylylsulfate: N-desulfoheparan sulfate sulfotransferase from arterial tissue
| Content Provider | Scilit |
|---|---|
| Author | Gohler, Dieter Niemann, Reinhard Buddecke, Eckhart |
| Copyright Year | 1984 |
| Description | Journal: Journal of Biological Inorganic Chemistry A 3'-phosphoadenylsulfate: N-desulfoheparan sulfate sulfotransferase (EC 2.8.2.12) was purified 450-fold from the microsomal fraction of calf arterial tissue and separated from 3'-phosphoadenylylsulfate:chondroitin sulfotransferase (EC 2.8.2.5) activity. The enzyme has optimal activity at neutral pH, requires divalent cations (Mn2+, Mg2+, Ca2+) for maximal activity and exhibits specificity towards N-desulfoheparan sulfate, N,O-desulfoheparan sulfate and oligosaccharides derived therefrom. N,O-desulfoheparan sulfate tetrasaccharides serve as acceptor substrates only if the nonreducing terminus is occupied by glucuronic acid (not iduronic acid). The N,O-desulfoheparan sulfate sulfotransferase transfers [35S]sulfate from 3'-phosphoadenylyl[35S]sulfate to the 2-amino groups and to the 6-hydroxy groups of glucosamine units of the acceptor substrates. The ratio of N/O-sulfation ranged between 3:1 and 2:1. O-[35S]Sulfated unsaturated disaccharides were obtained from enzymatically labelled [35S]N-desulfoheparan sulfate by heparitinase degradation and subsequent deamination. Evidence for the O-sulfation at C-6 of the glucosamine units was provided by isolation of anhydromannose [35S]monosulfate, which was formed from uronosylanhydromannose [35S]monosulfate by beta-glucuronidase treatment. An N-desulfo-N-[1-14C]lacetylheparan sulfate deacetylase activity was copurified with the N-desulfoheparan sulfate sulfotransferase. |
| Ending Page | 308 |
| Starting Page | 301 |
| ISSN | 2573508X |
| e-ISSN | 14321327 |
| DOI | 10.1111/j.1432-1033.1984.tb07915.x |
| Journal | Journal of Biological Inorganic Chemistry |
| Issue Number | 2 |
| Volume Number | 138 |
| Language | English |
| Publisher | Wiley-Blackwell |
| Publisher Date | 1984-01-01 |
| Access Restriction | Open |
| Subject Keyword | Journal: Journal of Biological Inorganic Chemistry Biochemistry and Molecular Biology Sulfate Sulfotransferase Desulfoheparan Sulfate |
| Content Type | Text |
| Resource Type | Article |
| Subject | Biochemistry Inorganic Chemistry |
