3-O-sulfation of heparan sulfate as a regular of protein function

项目摘要/摘要 肝素是肥大细胞产生的一种高度硫酸盐化的碳水化合物，被广泛用作抗凝剂。 预防深静脉血栓、肺血栓和动脉血栓形成。硫酸乙酰肝素 (HS)在结构上与肝素相关，装饰着几乎所有人类细胞的表面。它会相互作用 含有多种蛋白质，包括趋化因子和细胞因子；凝血因子；生长 与补体激活、细胞黏附和信号传递有关的因子和形态因子、蛋白质 蛋白质。HS与人类疾病有关，如血管疾病、炎症、 感染和神经退化。据推测，HS通过以下能力对信息进行编码 以上下文相关的方式招募蛋白质。尽管“HS-硫酸盐密码”假说是 从概念上讲，人们对HS结合蛋白的配体特异性和 HS生物合成酶的底物特异性。因此，目前尚不清楚HS的亚型是否 生物合成酶可以产生独特的表位，在细胞环境中可以招募特定的HS- 结合蛋白。了解HS约束的特殊性也提供了发展的机会 治疗方式。 在这个项目中，将开发化学和化学-酶方法，使其 可以制备具有和不具有3-O-硫酸盐的生物学相关HS表位的聚焦文库 半个月。这些化合物将用于开发HS-微阵列，以建立配体需求 HS结合蛋白。感兴趣的化合物将进一步探索它们干扰 内皮细胞的结合和细胞激活。我们将重点研究血液中的HS结合蛋白 凝血、炎症和血管疾病。已建立的结构-活性关系是 预计将提供治疗用的先导化合物。HS-微阵列也将被用作一般 发现和鉴定需要3-O-硫酸盐的HS结合蛋白的平台。此外， 合成化合物将用于确定各种3-O-磺基转移酶的底物要求。 总的来说，我们的研究将解决3-O-磺基转移酶的异构体是否可以 创造独特的表位，可以招募特定的HS结合蛋白，以介导各种生物 流程。 1

Project summary/abstract Heparin is a highly sulfated carbohydrate produced by mast cells that is widely used as an anticoagulant to prevent deep vein thrombosis, pulmonary embolism and arterial thromboembolism. Heparan sulfate (HS) is structurally related to heparin and decorates the surface of all most all human cells. It interacts with a multitude of proteins including chemokines and cytokines; blood coagulation factors; growth factors and morphogens, proteins involved in complement activation, and cell adhesion and signaling proteins. HS has been implicated in human diseases such as vascular diseases, inflammation, infections and neurodegeneration. It has been postulated that HS encodes information by an ability to recruit proteins in a context-dependent manner. Although the “HS-sulfate code” hypothesis is conceptually appealing, little is known about the ligand specificities of HS binding proteins and the substrate specificities of HS-biosynthetic enzymes. As a result, it is not known whether isoforms of HS biosynthetic enzymes can create distinctive epitopes that in a cellular context can recruit specific HS- binding proteins. An understanding of the specificities of HS binding also offers opportunities to develop therapeutic modalities. In this program, chemical and chemo-enzymatic methodologies will be developed that will make it possible to prepare a focused library of biologically relevant HS epitopes with and without a 3-O-sulfate moiety. These compounds will be used to develop an HS-microarray to establish ligand requirement of HS-binding proteins. Compounds of interest will be further explored for their ability to interfere in the binding and cellular activation of endothelial cells. We will focus on HS-binding proteins involved in blood coagulation, inflammation and vascular disease. The established structure-activity relationships are expected to provide therapeutic lead compounds. The HS-microarray will also be used as a general platform to discover and characterize HS-binding proteins requiring a 3-O-sulfate. In addition, the synthetic compounds will be used to define substrate requirements of the various 3-O-sulfotransferases. Collectively, our studies will address the question of whether isoforms of 3-O-sulfotransferases can create distinctive epitopes that can recruit specific HS-binding proteins, to mediate various biological processes. 1