HSulf-2, an extracellular endoglucosamine-6-sulfatase, selectively mobilizes heparin-bound growth factors and chemokines: effects on VEGF, FGF-1, and SDF-1.
HSulf-2, an extracellular endoglucosamine-6-sulfatase, selectively mobilizes heparin-bound growth factors and chemokines: effects on VEGF, FGF-1, and SDF-1.
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HSULF-2是一种细胞外内核葡萄糖-6-硫酸蛋白酶，有选择地动员了肝素结合的生长因子和趋化因子：对VEGF，FGF-1和SDF-1的影响。
DOI：
10.1186/1471-2091-7-2
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发表时间：
2006-01-17
期刊：
影响因子：
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通讯作者：
Rosen, Steven D
中科院分区：
文献类型：
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作者：
Uchimura, Kenji;Morimoto-Tomita, Megumi;Bistrup, Annette;Li, Jessica;Lyon, Malcolm;Gallagher, John;Werb, Zena;Rosen, Steven D
肝素/硫酸乙酰肝素 (HS) 蛋白多糖存在于细胞外基质 (ECM) 和细胞表面。大量证据表明，肝素和硫酸乙酰肝素蛋白聚糖 (HSPG) 与多种蛋白质配体相互作用，包括成纤维细胞生长因子、血管内皮生长因子 (VEGF)、细胞因子和趋化因子。这些相互作用高度依赖于糖胺聚糖链内硫酸化修饰的模式。我们之前克隆了编码新型人内硫酸酯酶 HSulf-2 的 cDNA，该酶可从完整肝素的亚区域中去除葡萄糖胺上的 6-O-硫酸基团。在这里，我们使用了重组 HSulf-2 和来自 MCF-7 乳腺癌细胞系条件培养基的天然酶。为了确定 HSulf-2 是否调节肝素结合因子和肝素之间的相互作用，我们开发了一种 ELISA，其中允许可溶性因子与固定化肝素结合。我们的结果表明，通过用 HSulf-2 预处理肝素，VEGF、FGF-1 和某些趋化因子（SDF-1 和 SLC）与固定化肝素的结合被消除或大大减少。此外，HSulf-2 将这些可溶性蛋白质从与肝素的结合中释放出来。来自 MCF-7 细胞的天然 Sulf-2 再现了所有这些活性。我们的结果验证了 Sulf-2 作为破译蛋白质配体与肝素/HSPG 相互作用中硫酸化要求的新工具，并扩大了该酶的潜在生物活性范围。
Heparin/heparan sulfate (HS) proteoglycans are found in the extracellular matrix (ECM) and on the cell surface. A considerable body of evidence has established that heparin and heparan sulfate proteoglycans (HSPGs) interact with numerous protein ligands including fibroblast growth factors, vascular endothelial growth factor (VEGF), cytokines, and chemokines. These interactions are highly dependent upon the pattern of sulfation modifications within the glycosaminoglycan chains. We previously cloned a cDNA encoding a novel human endosulfatase, HSulf-2, which removes 6-O-sulfate groups on glucosamine from subregions of intact heparin. Here, we have employed both recombinant HSulf-2 and the native enzyme from conditioned medium of the MCF-7-breast carcinoma cell line. To determine whether HSulf-2 modulates the interactions between heparin-binding factors and heparin, we developed an ELISA, in which soluble factors were allowed to bind to immobilized heparin. Our results show that the binding of VEGF, FGF-1, and certain chemokines (SDF-1 and SLC) to immobilized heparin was abolished or greatly diminished by pre-treating the heparin with HSulf-2. Furthermore, HSulf-2 released these soluble proteins from their association with heparin. Native Sulf-2 from MCF-7 cells reproduced all of these activities. Our results validate Sulf-2 as a new tool for deciphering the sulfation requirements in the interaction of protein ligands with heparin/HSPGs and expand the range of potential biological activities of this enzyme.