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Tykesson E, Persson A, Mani K, et al. Assays for Evaluation of Substrates for and Inhibitors of β-1,4-Galactosyltransferase 7. Methods Mol Biol. 2022;2303:477-486doi: 10.1007/978-1-0716-1398-6_38.
Tykesson, E., Persson, A., Mani, K., & Ellervik, U. (2022). Assays for Evaluation of Substrates for and Inhibitors of β-1,4-Galactosyltransferase 7. Methods in molecular biology (Clifton, N.J.), 2303477-486. https://doi.org/10.1007/978-1-0716-1398-6_38
Tykesson, Emil, et al. "Assays for Evaluation of Substrates for and Inhibitors of β-1,4-Galactosyltransferase 7." Methods in molecular biology (Clifton, N.J.) vol. 2303 (2022): 477-486. doi: https://doi.org/10.1007/978-1-0716-1398-6_38
Tykesson E, Persson A, Mani K, Ellervik U. Assays for Evaluation of Substrates for and Inhibitors of β-1,4-Galactosyltransferase 7. Methods Mol Biol. 2022;2303:477-486. doi: 10.1007/978-1-0716-1398-6_38. PMID: 34626402.
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Methods Mol Biol. 2022;2303:477-486. doi: 10.1007/978-1-0716-1398-6_38.

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

Methods in molecular biology (Clifton, N.J.)

Emil Tykesson, Andrea Persson, Katrin Mani, Ulf Ellervik

Affiliations

  1. Department of Experimental Medical Science, Division of Neuroscience, Glycobiology Group, Lund University, Lund, Sweden.
  2. Department of Laboratory Medicine, Institute of Biomedicine, Sahlgrenska Academy at the University of Gothenburg, Gothenburg, Sweden.
  3. Department of Experimental Medical Science, Division of Neuroscience, Glycobiology Group, Lund University, Lund, Sweden. [email protected].
  4. Department of Chemistry, Lund University, Lund, Sweden.

PMID: 34626402 DOI: 10.1007/978-1-0716-1398-6_38

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 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.

© 2022. Springer Science+Business Media, LLC, part of Springer Nature.

Keywords: Glycosaminoglycan biosynthesis; Inhibitor; Substrate; Xylosides; β-1,4-Galactosyltransferase 7

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