Heparan sulfate (HS) is a heterogeneous, extracellular glycan that interacts with proteins and other molecules affecting many biological processes. The specific binding motifs of HS interactions are of interest, but have not been extensively characterized. Glycan microarrays are valuable tools that can be used to probe the interactions between glycans and their ligands while relying on relatively small amounts of samples. Recently, chemoenzymatic synthesis of HS has been employed to produce specific HS structures that can otherwise be difficult to produce. In this study, a microarray of diverse chemoenzymatically synthesized HS structures was developed and HS interactions were characterized. Fluorescently labeled antithrombin III (AT) and fibroblast growth factor-2 (FGF2) were screened against 95 different HS structures under three different printing concentrations to confirm the utility of this microarray. Specific sulfation patterns were found to be important for binding to these proteins and results are consistent with previous specificity studies. Furthermore, the binding affinities (K_D,surf) of AT and FGF2 to multiple HS structures were determined using a microarray technique and is consistent with previous reports. Lastly, the 95-compound HS microarray was used to determine the distinct binding profiles for interleukin 12 and platelet factor 4. This technique is ideal for rapid expansion and will be pivotal to the high-throughput characterization of biologically important structure/function relationships.

中文翻译：

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硫酸乙酰肝素微阵列的构建用于研究特定糖序列与蛋白质的结合

硫酸乙酰肝素 (HS) 是一种异质的细胞外聚糖，可与影响许多生物过程的蛋白质和其他分子相互作用。HS 相互作用的特异性结合基序是令人感兴趣的，但尚未得到广泛的表征。聚糖微阵列是一种有价值的工具，可用于探测聚糖与其配体之间的相互作用，同时依赖于相对少量的样品。最近，HS 的化学酶合成已被用于生产可能难以生产的特定 HS 结构。在这项研究中，开发了多种化学酶合成的 HS 结构的微阵列，并对 HS 相互作用进行了表征。在三种不同的印刷浓度下针对 95 种不同的 HS 结构筛选荧光标记的抗凝血酶 III (AT) 和成纤维细胞生长因子-2 (FGF2)，以确认该微阵列的实用性。发现特定的硫酸化模式对于与这些蛋白质的结合很重要，结果与先前的特异性研究一致。此外，结合亲和力（K使用微阵列技术确定 AT 和 FGF2 对多个 HS 结构的_{D,surf ) 与先前的报道一致。}最后，使用 95 种化合物的 HS 微阵列来确定白细胞介素 12 和血小板因子 4 的不同结合谱。该技术非常适合快速扩增，对于生物学上重要的结构/功能关系的高通量表征至关重要。