Enzymatic modification of heparan sulfate on a biochip promotes its interaction with antithrombin III
Enzymatic modification of heparan sulfate on a biochip promotes its interaction with antithrombin III
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DOI：
10.1006/bbrc.2000.3453
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发表时间：
2000-09-16
影响因子：
3.1
通讯作者：
Linhardt, RJ
中科院分区：
文献类型：
--
作者：
Hernaiz, M;Liu, J;Linhardt, RJ
在还原端生物素化的硫酸肝素糖胺聚糖链与链霉亲和素包被的生物芯片结合，表面等离子体共振光谱显示，当它流过含有硫酸肝素的表面时，与抗凝血酶III （ATIII）具有低亲和力相互作用。在3'-磷酸腺苷5'-磷酸硫酸酯（PAPS）存在下，用3- o -硫代转移酶异构体1 （3- ost -1）对同一表面进行酶修饰后，ATIII以高亲和力紧密结合。3-OST-1酶参与硫酸肝素的生物合成，并在这种糖胺聚糖中引入一个关键的s - o -磺基，提供合适的五糖序列，能够与ATIII高亲和力结合。本实验展示了利用生物合成酶对结合在生物芯片上的糖胺聚糖进行特异性结构修饰，为快速筛选糖胺聚糖-蛋白质相互作用提供了一种新的方法。(C) 2000年学术出版社。
A heparan sulfate glycosaminoglycan chain, biotinylated at its reducing-end, was bound to a streptavidin-coated biochip, Surface plasmon resonance spectroscopy showed a low affinity interaction with antithrombin III (ATIII) when it was flowed over a surface containing heparan sulfate. ATIII bound tightly with high affinity when the same surface was enzymatically modified to using 3-O-sulfotransferase isoform 1 (3-OST-1) in the presence of 3'-phosphoadenosine 5'-phosphosulfate (PAPS). The 3-OST-1 enzyme is involved in heparan sulfate biosynthesis and introduces a critical S-O-sulfo group into this glycosaminoglycan affording the appropriate pentasaccharide sequence capable of high affinity binding to ATIII. This experiment demonstrates the specific structural modification of a glycosaminoglycan bound to a biochip using a biosynthetic enzyme, suggesting a new approach to rapid screening glycosaminoglycan-protein interactions. (C) 2000 Academic Press.