Deciphering mode of action of heparanase using structurally defined oligosaccharides.
Heparan sulfate (HS) is a highly sulfated polysaccharide that serves many biological functions, including regulating cell growth and inflammatory responses as well as the blood coagulation process. Heparanase is an enzyme that cleaves HS and is known to display a variety of pathophysiological effects in cancer, diabetes, and Alzheimer disease. The link between heparanase and diseases is a result of its selective cleavage of HS, which releases smaller HS fragments to enhance cell proliferation, migration, and invasion. Despite its importance in pathological diseases, the structural cues in HS that direct heparanase cleavage and the steps of HS depolymerization remain unknown. Here, we sought to probe the substrate specificity of heparanase using a series of structurally defined oligosaccharide substrates. The sites of heparanase cleavage on the oligosaccharide substrates were determined by mass spectrometry and gel permeation chromatography. We discovered that heparanase cleaves the linkage of glucuronic acid linked to glucosamine carrying 6-O-sulfo groups. Furthermore, our findings suggest that heparanase displays different cleavage modes by recognizing the structures of the nonreducing ends of the substrates. Our results deepen the understanding of the action mode of heparanase.
Division of Chemical Biology and Medicinal Chemistry, Eshelman School of Pharmacy, University of North Carolina, Chapel Hill, North Carolina 27599, USA.
Sherket
S
Peterson
Jian
J
Liu
D000544:Alzheimer Disease / Q000235:genetics / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D005944:Glucosamine / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D020723:Glucuronic Acid / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D005966:Glucuronidase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006497:Heparan Sulfate / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008956:Models, Chemical; D009844:Oligosaccharides / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D018411:Spodoptera; D000099175:Heparanase
D005944:Glucosamine; D020723:Glucuronic Acid; D005966:Glucuronidase; D006497:Heparan Sulfate; D009844:Oligosaccharides; D000099175:Heparanase