Measuring the activities of the Sulfs: two novel heparin/heparan sulfate endosulfatases.
Sulfatases hydrolyze sulfate esters on a variety of molecules including glycosaminoglycans, sulfoglycolipids, and cytosolic steroids. These enzymes are found in a wide range of organisms with their basic enzymatic mechanisms broadly conserved. In mammals, many of the sulfatases localize in the lysosome and exhibit enzymatic activity on a small aryl substrate such as 4-methylumbelliferyl sulfate (4-MUS). They are known as arylsulfatases. Sulf-1 and Sulf-2 have been cloned and identified as sulfatases that release sulfate groups on the C-6 position of GlcNAc residue from an internal subdomain in intact heparin. Hence, these enzymes are endosulfatases. The Sulfs are secreted in an active form into conditioned medium of transfected Chinese hamster ovary (CHO) cells. In this chapter, arylsulfatase and endoglucosamine-6-sulfatase assays for the Sulfs are described. A solid-phase binding assay is also detailed, which allows investigation of the ability of the Sulfs to modulate the interaction of heparin-binding proteins with immobilized heparin. The example illustrated is vascular endothelial growth factor (VEGF). This assay is projected to be very useful in the investigation of the biological functions of the Sulfs.
Uchimura
Kenji
K
Department of Anatomy, Program in Immunology, UCSF, USA.
Morimoto-Tomita
Megumi
M
Rosen
Steven D
SD
D001192:Arylsulfatases / Q000737:chemistry; D001667:Binding, Competitive; D001681:Biological Assay / Q000379:methods; D002478:Cells, Cultured; D006497:Heparan Sulfate / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D013379:Substrate Specificity; D013429:Sulfatases / Q000737:chemistry* / Q000378:metabolism; D015238:Sulfotransferases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D042461:Vascular Endothelial Growth Factor A / Q000737:chemistry
D001192:Arylsulfatases; D006497:Heparan Sulfate; D013429:Sulfatases; D015238:Sulfotransferases; D042461:Vascular Endothelial Growth Factor A; C470791:SULF1 protein, human; C470792:SULF2 protein, human; C021346:heparan sulfate sulfatase