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Assays for Evaluation of Substrates for and Inhibitors of β-1,4-Galactosyltransferase 7

Tykesson, Emil LU orcid ; Persson, Andrea LU ; Mani, Katrin LU orcid and Ellervik, Ulf LU orcid (2022) In Methods in Molecular Biology 2303. p.477-486
Abstract

β-1,4-Galactosyltransferase 7 (β4GalT7) is a key enzyme in the synthesis of two classes of glycosaminoglycans (GAG), i.e., heparan sulfate (HS) and chondroitin/dermatan sulfate (CS/DS). GAG chains are linear polysaccharides of alternating hexuronic acid and N-acetylhexosamine residues, commonly linked to core proteins to form proteoglycans with important roles in the regulation of a range of biological processes. The biosynthesis of GAGs is initiated by xylosylation of a serine residue of the core protein followed by galactosylation, catalyzed by β4GalT7. The biosynthesis can also be initiated by xylosides carrying hydrophobic aglycons, such as 2-naphthyl β-D-xylopyranoside. We have cloned and expressed β4GalT7, and designed a cell-free... (More)

β-1,4-Galactosyltransferase 7 (β4GalT7) is a key enzyme in the synthesis of two classes of glycosaminoglycans (GAG), i.e., heparan sulfate (HS) and chondroitin/dermatan sulfate (CS/DS). GAG chains are linear polysaccharides of alternating hexuronic acid and N-acetylhexosamine residues, commonly linked to core proteins to form proteoglycans with important roles in the regulation of a range of biological processes. The biosynthesis of GAGs is initiated by xylosylation of a serine residue of the core protein followed by galactosylation, catalyzed by β4GalT7. The biosynthesis can also be initiated by xylosides carrying hydrophobic aglycons, such as 2-naphthyl β-D-xylopyranoside. We have cloned and expressed β4GalT7, and designed a cell-free assay to measure the activity of this enzyme. The assay employs a 96-well plate format for high throughput. In this chapter, we describe the cloning, expression, and purification of β4GalT7, as well as assays proposed for development of substrates for GAG priming and for investigating inhibitors of β4GalT7.

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Please use this url to cite or link to this publication:
author
; ; and
organization
publishing date
type
Chapter in Book/Report/Conference proceeding
publication status
published
subject
keywords
Glycosaminoglycan biosynthesis, Inhibitor, Substrate, Xylosides, β-1,4-Galactosyltransferase 7
host publication
Methods in Molecular Biology
series title
Methods in Molecular Biology
volume
2303
pages
10 pages
publisher
Humana Press
external identifiers
  • pmid:34626402
  • scopus:85117133446
ISSN
1940-6029
1064-3745
DOI
10.1007/978-1-0716-1398-6_38
language
English
LU publication?
yes
additional info
Publisher Copyright: © 2022, Springer Science+Business Media, LLC, part of Springer Nature.
id
59af3c08-c59c-4bef-a08d-59f9425fe0af
date added to LUP
2021-11-01 13:43:43
date last changed
2024-06-01 19:03:22
@inbook{59af3c08-c59c-4bef-a08d-59f9425fe0af,
  abstract     = {{<p>β-1,4-Galactosyltransferase 7 (β4GalT7) is a key enzyme in the synthesis of two classes of glycosaminoglycans (GAG), i.e., heparan sulfate (HS) and chondroitin/dermatan sulfate (CS/DS). GAG chains are linear polysaccharides of alternating hexuronic acid and N-acetylhexosamine residues, commonly linked to core proteins to form proteoglycans with important roles in the regulation of a range of biological processes. The biosynthesis of GAGs is initiated by xylosylation of a serine residue of the core protein followed by galactosylation, catalyzed by β4GalT7. The biosynthesis can also be initiated by xylosides carrying hydrophobic aglycons, such as 2-naphthyl β-D-xylopyranoside. We have cloned and expressed β4GalT7, and designed a cell-free assay to measure the activity of this enzyme. The assay employs a 96-well plate format for high throughput. In this chapter, we describe the cloning, expression, and purification of β4GalT7, as well as assays proposed for development of substrates for GAG priming and for investigating inhibitors of β4GalT7.</p>}},
  author       = {{Tykesson, Emil and Persson, Andrea and Mani, Katrin and Ellervik, Ulf}},
  booktitle    = {{Methods in Molecular Biology}},
  issn         = {{1940-6029}},
  keywords     = {{Glycosaminoglycan biosynthesis; Inhibitor; Substrate; Xylosides; β-1,4-Galactosyltransferase 7}},
  language     = {{eng}},
  pages        = {{477--486}},
  publisher    = {{Humana Press}},
  series       = {{Methods in Molecular Biology}},
  title        = {{Assays for Evaluation of Substrates for and Inhibitors of β-1,4-Galactosyltransferase 7}},
  url          = {{http://dx.doi.org/10.1007/978-1-0716-1398-6_38}},
  doi          = {{10.1007/978-1-0716-1398-6_38}},
  volume       = {{2303}},
  year         = {{2022}},
}