Substrate specificities of mouse heparan sulphate glucosaminyl 6-O-sulphotransferases
(2003) In Biochemical Journal 372(Pt 2). p.80-371- Abstract
Glycosaminoglycan heparan sulphate interacts with a variety of proteins, such as growth factors, cytokines, enzymes and inhibitors and, thus, influences cellular functions, including adhesion, motility, differentiation and morphogenesis. The interactions generally involve saccharide domains in heparan sulphate chains, with precisely located O-sulphate groups. The 6-O-sulphate groups on glucosamine units, supposed to be involved in various interactions of functional importance, occur in different structural contexts. Three isoforms of the glucosaminyl 6-O-sulphotransferase (6-OST) have been cloned and characterized [H. Habuchi, M. Tanaka, O. Habuchi, K. Yoshida, H. Suzuki, K. Ban and K. Kimata (2000) J. Biol. Chem. 275, 2859-2868]. We... (More)
Glycosaminoglycan heparan sulphate interacts with a variety of proteins, such as growth factors, cytokines, enzymes and inhibitors and, thus, influences cellular functions, including adhesion, motility, differentiation and morphogenesis. The interactions generally involve saccharide domains in heparan sulphate chains, with precisely located O-sulphate groups. The 6-O-sulphate groups on glucosamine units, supposed to be involved in various interactions of functional importance, occur in different structural contexts. Three isoforms of the glucosaminyl 6-O-sulphotransferase (6-OST) have been cloned and characterized [H. Habuchi, M. Tanaka, O. Habuchi, K. Yoshida, H. Suzuki, K. Ban and K. Kimata (2000) J. Biol. Chem. 275, 2859-2868]. We have studied the substrate specificities of the recombinant enzymes using various O-desulphated poly- and oligo-saccharides as substrates, and using adenosine 3'-phosphate 5'-phospho[(35)S]sulphate as sulphate donor. All three enzymes catalyse 6-O-sulphation of both -GlcA-GlcNS- and -IdoA-GlcNS- (where GlcA represents D-glucuronic acid, NS the N-sulphate group and IdoA the L-iduronic acid) sequences, with preference for IdoA-containing targets, with or without 2-O-sulphate substituents. 6-OST1 showed relatively higher activity towards target sequences lacking 2-O-sulphate, e.g. the -GlcA-GlcNS- disaccharide unit. Sulphation of such non-O-sulphated acceptor sequences was generally favoured at low acceptor polysaccharide concentrations. Experiments using partially O-desulphated antithrombin-binding oligosaccharide as the acceptor revealed 6-O-sulphation of N-acetylated as well as 3-O-sulphated glucosamine residues with each of the three 6-OSTs. We conclude that the three 6-OSTs have qualitatively similar substrate specificities, with minor differences in target preference.
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- author
- Smeds, Emanuel LU ; Habuchi, Hiroko ; Do, Anh-Tri ; Hjertson, Eva ; Grundberg, Helena ; Kimata, Koji ; Lindahl, Ulf and Kusche-Gullberg, Marion
- publishing date
- 2003
- type
- Contribution to journal
- publication status
- published
- keywords
- Animals, COS Cells/enzymology, Carbohydrate Sequence, Cattle, Chlorocebus aethiops, Chromatography, High Pressure Liquid, Glucosamine/metabolism, Heparan Sulfate Proteoglycans/metabolism, Intestinal Mucosa/metabolism, Lung/metabolism, Mice, Molecular Sequence Data, Oligosaccharides/chemistry, Protein Isoforms, Recombinant Fusion Proteins/genetics, Substrate Specificity, Sulfotransferases/chemistry, Swine
- in
- Biochemical Journal
- volume
- 372
- issue
- Pt 2
- pages
- 80 - 371
- publisher
- Portland Press
- external identifiers
-
- scopus:0038677010
- pmid:12611590
- ISSN
- 0264-6021
- DOI
- 10.1042/BJ20021666
- language
- English
- LU publication?
- no
- id
- 587fd9b9-c03d-4ea3-8ad3-a33b05b35f95
- date added to LUP
- 2021-07-01 16:49:27
- date last changed
- 2024-03-08 15:26:25
@article{587fd9b9-c03d-4ea3-8ad3-a33b05b35f95,
abstract = {{<p>Glycosaminoglycan heparan sulphate interacts with a variety of proteins, such as growth factors, cytokines, enzymes and inhibitors and, thus, influences cellular functions, including adhesion, motility, differentiation and morphogenesis. The interactions generally involve saccharide domains in heparan sulphate chains, with precisely located O-sulphate groups. The 6-O-sulphate groups on glucosamine units, supposed to be involved in various interactions of functional importance, occur in different structural contexts. Three isoforms of the glucosaminyl 6-O-sulphotransferase (6-OST) have been cloned and characterized [H. Habuchi, M. Tanaka, O. Habuchi, K. Yoshida, H. Suzuki, K. Ban and K. Kimata (2000) J. Biol. Chem. 275, 2859-2868]. We have studied the substrate specificities of the recombinant enzymes using various O-desulphated poly- and oligo-saccharides as substrates, and using adenosine 3'-phosphate 5'-phospho[(35)S]sulphate as sulphate donor. All three enzymes catalyse 6-O-sulphation of both -GlcA-GlcNS- and -IdoA-GlcNS- (where GlcA represents D-glucuronic acid, NS the N-sulphate group and IdoA the L-iduronic acid) sequences, with preference for IdoA-containing targets, with or without 2-O-sulphate substituents. 6-OST1 showed relatively higher activity towards target sequences lacking 2-O-sulphate, e.g. the -GlcA-GlcNS- disaccharide unit. Sulphation of such non-O-sulphated acceptor sequences was generally favoured at low acceptor polysaccharide concentrations. Experiments using partially O-desulphated antithrombin-binding oligosaccharide as the acceptor revealed 6-O-sulphation of N-acetylated as well as 3-O-sulphated glucosamine residues with each of the three 6-OSTs. We conclude that the three 6-OSTs have qualitatively similar substrate specificities, with minor differences in target preference.</p>}},
author = {{Smeds, Emanuel and Habuchi, Hiroko and Do, Anh-Tri and Hjertson, Eva and Grundberg, Helena and Kimata, Koji and Lindahl, Ulf and Kusche-Gullberg, Marion}},
issn = {{0264-6021}},
keywords = {{Animals; COS Cells/enzymology; Carbohydrate Sequence; Cattle; Chlorocebus aethiops; Chromatography, High Pressure Liquid; Glucosamine/metabolism; Heparan Sulfate Proteoglycans/metabolism; Intestinal Mucosa/metabolism; Lung/metabolism; Mice; Molecular Sequence Data; Oligosaccharides/chemistry; Protein Isoforms; Recombinant Fusion Proteins/genetics; Substrate Specificity; Sulfotransferases/chemistry; Swine}},
language = {{eng}},
number = {{Pt 2}},
pages = {{80--371}},
publisher = {{Portland Press}},
series = {{Biochemical Journal}},
title = {{Substrate specificities of mouse heparan sulphate glucosaminyl 6-O-sulphotransferases}},
url = {{http://dx.doi.org/10.1042/BJ20021666}},
doi = {{10.1042/BJ20021666}},
volume = {{372}},
year = {{2003}},
}