Trisaccharide Sulfate and Its Sulfonamide as an Effective Substrate and Inhibitor of Human Endo-O-sulfatase-1
Trisaccharide Sulfate and Its Sulfonamide as an Effective Substrate and Inhibitor of Human Endo-O-sulfatase-1
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DOI：
10.1021/jacs.0c00005
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发表时间：
2020-03-18
影响因子：
15
通讯作者：
Hung, Shang-Cheng
中科院分区：
文献类型：
--
作者：
Chiu, Li-Ting;Sabbavarapu, Narayana Murthy;Hung, Shang-Cheng
人内切-O-硫酸酯酶（Sulf-1 和 Sulf-2）是细胞外硫酸乙酰肝素蛋白多糖 (HSPG) 特异性的 6-O-内切硫酸酯酶，可通过硫酸乙酰肝素 (HS)-蛋白质相互作用调节多种细胞信号转导事件，并与骨关节炎的发病相关。这些内切-O-硫酸酯酶被转运到细胞表面，从 HSPG 高度硫酸化亚结构域的内部 D-葡萄糖胺残基中释放 6-硫酸基团。在这项研究中，通过化学合成的多种具有不同链长和N-和O-硫酸化模式的HS寡糖在竞争测定中使用荧光底物4-甲基伞形基硫酸盐(4-MUS)系统地研究了人Sulf-1的底物特异性。硫酸三糖 IdoA2S-GlcNS6S-IdoA2S 被发现是 Sulf-1 的最小尺寸底物，用磺酰胺基序取代 D-葡萄糖胺单元 6-O 位的硫酸基团可有效抑制 Sulf-1 活性，IC50 = 0.53 μM，K-i = 0.36 μM，K-D = 12 纳米。
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 mu M, K-i = 0.36 mu M, and K-D = 12 nM.