Enzymatic synthesis of heparin related polysaccharides on sensor chips:: Rapid screening of heparin-protein interactions
Enzymatic synthesis of heparin related polysaccharides on sensor chips:: Rapid screening of heparin-protein interactions
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DOI：
10.1016/j.bbrc.2005.11.051
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发表时间：
2006-01-13
影响因子：
3.1
通讯作者：
Linhardt, RJ
中科院分区：
文献类型：
--
作者：
Muñoz, E;Xu, D;Linhardt, RJ
肝素（HP）和硫酸乙酰肝素（HS）的生物学作用主要通过它们与蛋白质的相互作用来介导。在目前的工作中，我们提供了一个快速的方法来筛选HP/HS-蛋白质的相互作用提供的结构数据的关键磺基参与结合。通过将生物素化的N-硫酸化K5多糖（N-磺基肝素原）固定在传感器芯片上，然后用参与HP/HS生物合成的酶选择性修饰该多糖，制备结构上与HP相关的多糖库。在传感器芯片表面上合成的多糖在糖醛酸和葡糖胺残基上存在的磺基的数量和位置上不同。表面等离子体共振用于测量该多糖库的每个成员与抗凝血酶III（ATIII）的相互作用，以提供该HP/HS-蛋白质相互作用所需的磺基的结构信息。这种方法被认为是广泛适用于HP/HS-蛋白质相互作用的构效关系（SAR）的研究。(c)2005年爱思唯尔公司All rights reserved.
The biological roles of heparin (HP) and heparan sulfate (HS) are mediated mainly through their interaction with proteins. In the present work, we provide a rapid method for screening HP/HS-protein interactions providing structural data on the key sulfo groups that participate in the binding. A library of polysaccharides structurally related to HP was prepared by immobilizing the biotinylated N-sulfated K5 polysaccharide (N-sulfoheparosan) on sensor chips followed by selective modification of this polysaccharide with enzymes that participate in HP/HS biosynthesis. The polysaccharides synthesized on the surface of the sensor chips differ in the number and position of sulfo groups present both on uronic acid and glucosamine residues. Surface plasmon resonance was used to measure the interaction of each member of this polysaccharide library with antithrombin III (ATIII), to afford structural information on sulfo groups required for this HP/HS-protein interaction. This method is viewed as widely applicable for the study of the structure-activity relationship (SAR) of HP/HS-protein interactions. (c) 2005 Elsevier Inc. All rights reserved.