Functional consequences of the subdomain organization of the sulfs.
Sulf-1 and Sulf-2 are novel extracellular sulfatases that act on internal glucosamine 6-O-sulfate modifications within heparan sulfate proteoglycans and regulate their interactions with various signaling molecules, including Wnt ligands. Although the Sulfs are multidomain proteins, there is limited information available about how the subdomains contribute to their enzymatic and signaling activities. In this study, we found that both human Sulfs were synthesized as prepro-enzymes and cleaved by a furin-type proteinase to form disulfide-bond linked heterodimers of 75- and 50-kDa subunits. The mature Sulfs were secreted into conditioned medium, as well as retained on the cell membrane. Although the catalytic center resides in the N-terminal 75-kDa subunit, the C-terminal 50-kDa subunit was indispensable for both arylsufatase and glucosamine 6-O-sulfate-endosulfatase activity. We found that the hydrophilic regions of the Sulfs were essential for endosulfatase activity but not for arylsulfatase activity. Using Edman sequencing, we identified furin-type proteinase cleavage sites in Sulf-1 and Sulf-2. Deletion of these sequences resulted in uncleavable forms of Sulfs. The uncleavable Sulfs retained enzymatic activity. However, they were unable to potentiate Wnt signaling, which may be due to their defective localization into lipid rafts on the plasma membrane.
Tang
Renhong
R
Department of Anatomy, University of California, San Francisco, California 94143, USA.
Rosen
Steven D
SD
D000595:Amino Acid Sequence; D002384:Catalysis; D002462:Cell Membrane / Q000378:metabolism; D004220:Disulfides / Q000737:chemistry; D045683:Furin / Q000737:chemistry; D006801:Humans; D008024:Ligands; D021962:Membrane Microdomains / Q000737:chemistry; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D017386:Sequence Homology, Amino Acid; D015398:Signal Transduction; D013429:Sulfatases; D015238:Sulfotransferases / Q000737:chemistry / Q000502:physiology*; D051153:Wnt Proteins / Q000378:metabolism
D004220:Disulfides; D008024:Ligands; D051153:Wnt Proteins; C470791:SULF1 protein, human; D015238:Sulfotransferases; C470792:SULF2 protein, human; D013429:Sulfatases; D045683:Furin