Trisaccharide Sulfate and Its Sulfonamide as an Effective Substrate and Inhibitor of Human Endo‑O‑sulfatase‑1

• Published in: Journal of the American Chemical Society Vol. 142; no. 11; pp. 5282 - 5292
• Main Authors: Chiu, Li-Ting, Sabbavarapu, Narayana Murthy, Lin, Wei-Chen, Fan, Chiao-Yuan, Wu, Chih-Chung, Cheng, Ting-Jen Rachel, Wong, Chi-Huey, Hung, Shang-Cheng
• Format: Journal Article
• Language: English
• Published: WASHINGTON American Chemical Society 18.03.2020; Amer Chemical Soc
• Subjects: Chemistry; Chemistry, Multidisciplinary; Enzyme Assays; Enzyme Inhibitors - chemical synthesis; Enzyme Inhibitors - chemistry; enzymes; glucosamine; heparan sulfate; Heparitin Sulfate - chemistry; Humans; inhibitory concentration 50; Kinetics; osteoarthritis; Physical Sciences; proteoglycans; Science & Technology; Substrate Specificity; Sulfatases - antagonists & inhibitors; Sulfatases - chemistry; sulfates; Sulfonamides - chemical synthesis; Sulfonamides - chemistry; Sulfotransferases - antagonists & inhibitors; Sulfotransferases - chemistry; synthesis; trisaccharides; Trisaccharides - chemical synthesis; Trisaccharides - chemistry; PROTEOGLYCANS; COMPLEX; SULF-2; SPECIFICITY; CHEMOENZYMATIC SYNTHESIS; GROWTH; HEPARAN-SULFATE; DEFICIENCY
• ISSN: 0002-7863; 1520-5126; 1520-5126
• DOI: 10.1021/jacs.0c00005

Human endo-O-sulfatases (Sulf-1 and Sulf-2) are extracellular heparan sulfate proteoglycan (HSPG)-specific 6-O-endosulfatases, which regulate a multitude of cell-signaling events through heparan sulfate (HS)–protein interactions and are associated with the onset of osteoarthritis. These endo-O-sulfatases are transported onto the cell surface to liberate the 6-sulfate groups from the internal d-glucosamine residues in the highly sulfated subdomains of HSPGs. In this study, a variety of HS oligosaccharides with different chain lengths and N- and O-sulfation patterns via chemical synthesis were systematically studied about the substrate specificity of human Sulf-1 employing the fluorogenic substrate 4-methylumbelliferyl sulfate (4-MUS) in a competition assay. The trisaccharide sulfate IdoA2S-GlcNS6S-IdoA2S was found to be the minimal-size substrate for Sulf-1, and substitution of the sulfate group at the 6-O position of the d-glucosamine unit with the sulfonamide motif effectively inhibited the Sulf-1 activity with IC50 = 0.53 μM, K i = 0.36 μM, and K D = 12 nM.