Title :

The Sulfs: expression, purification, and substrate specificity

Author(s) :

Uchimura, Kenji [Auteur]
Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 [UGSF]

Journal title :

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

Abbreviated title :

Methods Mol. Biol.

Volume number :

1229

Pages :

401-412

Publication date :

2015

ISSN :

1940-6029

English keyword(s) :

Brain
Disaccharides
Humans
Heparitin Sulfate
Substrate Specificity
Sulfatases
Immunoblotting
Enzyme Assays
Recombinant Proteins
Epitopes
Plaque, Amyloid
Animals
Transfection
HEK293 Cells
Mice

HAL domain(s) :

Chimie/Chimie théorique et/ou physique

English abstract : [en]

Sulf-1 and Sulf-2 are endo-acting extracellular sulfatases. The Sulfs liberate 6-O sulfate groups, mainly from N, 6-O, and 2-O trisulfated disaccharides of heparan sulfate (HS)/heparin chains. The Sulfs have been shown to ...
Show more >Sulf-1 and Sulf-2 are endo-acting extracellular sulfatases. The Sulfs liberate 6-O sulfate groups, mainly from N, 6-O, and 2-O trisulfated disaccharides of heparan sulfate (HS)/heparin chains. The Sulfs have been shown to modulate the interaction of a number of protein ligands including growth factors and morphogens with HS/heparin and thus regulate the signaling of these ligands. They also play important roles in development and are dysregulated in many cancers. The establishment of the expression of the Sulfs and methods of assaying them has been desirable to investigate these enzymes. In this chapter, methods to express and purify recombinant Sulfs and to analyze HS structures in an extracellular fraction of HSulf-transfected HEK293 cells are described. The application of these enzymes for ex vivo degradation of an anti-HS epitope accumulated in the brain of a neurodegenerative disease model mouse is also described.Show less >

Language :

Anglais

Administrative institution(s) :

CNRS
Université de Lille

Collections :

• Unité de Glycobiologie Structurale et Fonctionnelle (UGSF) - UMR 8576

Submission date :

2020-02-12T15:11:37Z