A Cluster Sequencing Strategy To Determine the Consensus Affinity Domains in Heparin for Its Binding to Specific Proteins
A Cluster Sequencing Strategy To Determine the Consensus Affinity Domains in Heparin for Its Binding to Specific Proteins
复制标题
用于确定肝素与特定蛋白质结合的共有亲和域的聚类测序策略
DOI：
10.1021/acs.analchem.2c03267
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发表时间：
2022-10-02
影响因子：
7.4
通讯作者：
Chi，Lianli
中科院分区：
文献类型：
--
作者：
Shi，Deling;Sheng，Anran;Chi，Lianli
糖胺聚糖（GAG）具有高负电荷，并且在生物学和药学上是重要的，因为它们的高电荷促进与许多蛋白质的强相互作用。由于GAG的固有异质性，含有某些共同结构域的多个寡糖通常可以与靶蛋白上的碱性氨基酸残基簇相互作用。许多GAG-蛋白质相互作用的特异性仍未被发现，因为关于相互作用的GAG的结构信息不足。在此，我们建立了一个集群测序策略，同时推导出所有的主要序列的亲和性GAG寡糖，导致定义的共识序列，它们共享对应于目标蛋白的特异性结合结构域。作为一个概念的证明，抗凝血酶III结合寡糖进行了检查，导致在一个七糖结构域含有完善的抗凝五糖序列。重复这种方法，发现了一个新的五糖结构域，对应于负责结合干扰素-γ（IFNγ）的肝素基序。我们的策略对于发现开发新的基于GAG的治疗剂所需的糖序列至关重要。
Glycosaminoglycans (GAGs) have high negative charge and are biologically and pharmaceutically important because their high charge promotes a strong interaction with many proteins. Due to the inherent heterogeneity of GAGs, multiple oligosaccharides, containing certain common domains, often can interact with clusters of basic amino acid residues on a target protein. The specificity of many GAG–protein interactions remains undiscovered since there is insufficient structural information on the interacting GAGs. Herein, we establish a cluster sequencing strategy to simultaneously deduce all major sequences of the affinity GAG oligosaccharides, leading to a definition of the consensus sequence they share that corresponds to the specific binding domain for the target protein. As a proof of concept, antithrombin III-binding oligosaccharides were examined, resulting in a heptasaccharide domain containing the well-established anticoagulant pentasaccharide sequence. Repeating this approach, a new pentasaccharide domain was discovered corresponding to the heparin motif responsible for binding interferon-γ (IFNγ). Our strategy is fundamentally important for the discovery of saccharide sequences needed in the development of novel GAG-based therapeutics.