Photoaffinity Probes for the Identification of Sequence-Specific Glycosaminoglycan-Binding Proteins,Journal of the American Chemical Society

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Photoaffinity Probes for the Identification of Sequence-Specific Glycosaminoglycan-Binding Proteins
Journal of the American Chemical Society ( IF 14.4 ) Pub Date : 2020-07-27 , DOI: 10.1021/jacs.0c06046
Amélie M Joffrin ₁ , Linda C Hsieh-Wilson ₁

Affiliation

Glycosaminoglycan (GAG)-protein interactions mediate critical physiological and pathological processes, such as neuronal plasticity, development, and viral invasion. However, mapping GAG-protein interaction networks is challenging as these interactions often require specific GAG sulfation patterns and involve transmembrane receptors or extracellular matrix-associated proteins. Here, we report the first GAG polysaccharide-based photoaffinity probes for the system-wide identification of GAG-binding proteins in living cells. A general platform for the modular, efficient assembly of various chondroitin sulfate (CS)-based photoaffinity probes was developed. Systematic evaluations led to benzophenone-containing probes that efficiently and selectively captured known CS-E-binding proteins in vitro and in cells. Importantly, the probes also enabled the identification of >50 new proteins from living neurons that interact with the neuroplasticity-relevant CS-E sulfation motif. Several candidates were independently validated and included membrane receptors important for axon guidance, innate immunity, synapse development, and synaptic plasticity. Overall, our studies provide a powerful approach for mapping GAG-protein interaction networks, revealing new potential functions for these polysaccharides and linking them to diseases such as Alzheimer's and autism.

中文翻译：

用于鉴定序列特异性糖胺聚糖结合蛋白的光亲和探针

糖胺聚糖 (GAG)-蛋白质相互作用介导关键的生理和病理过程，如神经元可塑性、发育和病毒入侵。然而，绘制 GAG-蛋白质相互作用网络具有挑战性，因为这些相互作用通常需要特定的 GAG 硫酸化模式并涉及跨膜受体或细胞外基质相关蛋白。在这里，我们报告了第一个基于 GAG 多糖的光亲和探针，用于全系统识别活细胞中的 GAG 结合蛋白。开发了用于模块化、高效组装各种基于硫酸软骨素 (CS) 的光亲和探针的通用平台。系统评估导致含二苯甲酮的探针在体外和细胞中有效和选择性地捕获已知的 CS-E 结合蛋白。重要的，这些探针还能够从与神经可塑性相关的 CS-E 硫酸化基序相互作用的活神经元中鉴定出超过 50 种新蛋白质。几个候选物经过独立验证，包括对轴突引导、先天免疫、突触发育和突触可塑性很重要的膜受体。总的来说，我们的研究为绘制 GAG-蛋白质相互作用网络提供了一种强大的方法，揭示了这些多糖的新潜在功能并将它们与阿尔茨海默氏症和自闭症等疾病联系起来。

更新日期：2020-07-27

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