Quail Sulf1 function requires asparagine-linked glycosylation.
The heparan sulfate endosulfatases Sulf1 and Sulf2 are cell-surface enzymes that control growth factor signaling through regulation of the 6-O-sulfation states of cell-surface and matrix heparan sulfate proteoglycans. Here, we report that quail Sulf1 (QSulf1) is an asparagine-linked glycosylated protein. Domain mapping studies in combination with a protein glycosylation prediction program identified multiple asparagine-linked glycosylation sites in the enzymatic and C-terminal domains. Glycosylation inhibitor studies revealed that glycosylation of QSulf1 is essential for its enzymatic activity, membrane targeting, and secretion. Furthermore, N-glycanase cleavage of asparagine-linked sites in native QSulf1 provided direct evidence that these N-linked glycosylation sites are specifically required for QSulf1 heparin binding and its 6-O-desulfation activity, revealing that N-linked glycosylation has a key role in the control of sulfatase enzymatic function.
Ambasta
Rashmi K
RK
Boston Biomedical Research Institute, 64 Grove Street, Watertown, MA 02472, USA.
Ai
Xingbin
X
Emerson
Charles P
CP
D000818:Animals; D001216:Asparagine / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000201:enzymology*; D015870:Gene Expression; D006031:Glycosylation; D006493:Heparin / Q000378:metabolism; D006497:Heparan Sulfate / Q000378:metabolism*; D006801:Humans; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase / Q000494:pharmacology; D011499:Protein Processing, Post-Translational / Q000187:drug effects / Q000502:physiology*; D017434:Protein Structure, Tertiary / Q000502:physiology; D011784:Quail / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins; D015398:Signal Transduction / Q000502:physiology*; D015238:Sulfotransferases / Q000235:genetics / Q000378:metabolism*
D001216:Asparagine; D006493:Heparin; D006497:Heparan Sulfate; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; D011994:Recombinant Proteins; D015238:Sulfotransferases