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The 3-O-sulfation of heparan sulfate modulates protein binding and lyase degradation [Chemistry]
Proceedings of the National Academy of Sciences of the United States of America ( IF 9.4 ) Pub Date : 2021-01-19 , DOI: 10.1073/pnas.2012935118
Pradeep Chopra 1 , Apoorva Joshi 1, 2 , Jiandong Wu 3 , Weigang Lu 1, 2 , Tejabhiram Yadavalli 4 , Margreet A Wolfert 1, 5 , Deepak Shukla 4, 6 , Joseph Zaia 3 , Geert-Jan Boons 2, 5, 7
Affiliation  

Humans express seven heparan sulfate (HS) 3-O-sulfotransferases that differ in substrate specificity and tissue expression. Although genetic studies have indicated that 3-O-sulfated HS modulates many biological processes, ligand requirements for proteins engaging with HS modified by 3-O-sulfate (3-OS) have been difficult to determine. In particular, the context in which the 3-OS group needs to be presented for binding is largely unknown. We describe herein a modular synthetic approach that can provide structurally diverse HS oligosaccharides with and without 3-OS. The methodology was employed to prepare 27 hexasaccharides that were printed as a glycan microarray to examine ligand requirements of a wide range of HS-binding proteins. The binding selectivity of antithrombin-III (AT-III) compared well with anti-Factor Xa activity supporting robustness of the array technology. Many of the other examined HS-binding proteins required an IdoA2S-GlcNS3S6S sequon for binding but exhibited variable dependence for the 2-OS and 6-OS moieties, and a GlcA or IdoA2S residue neighboring the central GlcNS3S. The HS oligosaccharides were also examined as inhibitors of cell entry by herpes simplex virus type 1, which, surprisingly, showed a lack of dependence of 3-OS, indicating that, instead of glycoprotein D (gD), they competitively bind to gB and gC. The compounds were also used to examine substrate specificities of heparin lyases, which are enzymes used for depolymerization of HS/heparin for sequence determination and production of therapeutic heparins. It was found that cleavage by lyase II is influenced by 3-OS, while digestion by lyase I is only affected by 2-OS. Lyase III exhibited sensitivity to both 3-OS and 2-OS.



中文翻译:

硫酸乙酰肝素的 3-O-硫酸化调节蛋白质结合和裂解酶降解 [化学]

人类表达七种在底物特异性和组织表达方面不同的硫酸乙酰肝素 (HS) 3 - O-磺基转移酶。尽管遗传研究表明 3 - O-硫酸化 HS 调节许多生物过程,但与 3- O修饰的 HS 结合的蛋白质的配体需求-硫酸盐(3-OS)一直难以确定。特别是,需要为绑定提供 3-OS 组的上下文在很大程度上是未知的。我们在本文中描述了一种模块化合成方法,该方法可以提供具有和不具有 3-OS 的结构多样的 HS 寡糖。该方法用于制备 27 种六糖,这些六糖被打印为聚糖微阵列,以检查各种 HS 结合蛋白的配体需求。抗凝血酶-III (AT-III) 的结合选择性与支持阵列技术稳健性的抗 Xa 因子活性相比较。许多其他检查的 HS 结合蛋白需要 IdoA2S-GlcNS3S6S 序列进行结合,但对 2-OS 和 6-OS 部分以及与中央 GlcNS3S 相邻的 GlcA 或 IdoA2S 残基表现出可变的依赖性。HS 寡糖也被检查为 1 型单纯疱疹病毒细胞进入的抑制剂,令人惊讶的是,它显示出缺乏对 3-OS 的依赖性,这表明它们竞争性结合 gB 和 gC 而不是糖蛋白 D (gD) . 这些化合物还用于检查肝素裂解酶的底物特异性,肝素裂解酶是用于解聚 HS/肝素的酶,用于序列测定和治疗性肝素的生产。发现裂解酶 II 的切割受 3-OS 的影响,而裂解酶 I 的消化仅受 2-OS 的影响。Lyase III 对 3-OS 和 2-OS 都表现出敏感性。这些化合物还用于检查肝素裂解酶的底物特异性,肝素裂解酶是用于解聚 HS/肝素的酶,用于序列测定和治疗性肝素的生产。发现裂解酶 II 的切割受 3-OS 的影响,而裂解酶 I 的消化仅受 2-OS 的影响。Lyase III 对 3-OS 和 2-OS 都表现出敏感性。这些化合物还用于检查肝素裂解酶的底物特异性,肝素裂解酶是用于解聚 HS/肝素的酶,用于序列测定和治疗性肝素的生产。发现裂解酶 II 的切割受 3-OS 的影响,而裂解酶 I 的消化仅受 2-OS 的影响。Lyase III 对 3-OS 和 2-OS 都表现出敏感性。

更新日期:2021-01-14
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