Hassan Abdullah A, Wozniak Jacob M, Vilen Zak, Li Weichao, Jadhav Appaso, Parker Christopher G, Huang Mia L
Department of Molecular Medicine, Scripps Research, 10550 N Torrey Pines Rd. La Jolla CA 92037 USA
Department of Chemistry, Scripps Research, 10550 N Torrey Pines Rd. La Jolla CA 92037 USA
RSC Chem Biol. 2022 Oct 18;3(12):1369-1374. doi: 10.1039/d2cb00176d. eCollection 2022 Nov 30.
Human milk oligosaccharides (HMOs) are a family of unconjugated soluble glycans found in human breast milk that exhibit a myriad of biological activity. While recent studies have uncovered numerous biological functions for HMOs (antimicrobial, anti-inflammatory & probiotic properties), the receptors and protein binding partners involved in these processes are not well characterized. This can be attributed largely in part to the low affinity and transient nature of soluble glycan-protein interactions, precluding the use of traditional characterization techniques to survey binding partners in live cells. Here, we present the use of synthetic photoactivatable HMO probes to capture, enrich and identify HMO protein targets in live cells using mass spectrometry-based chemoproteomics. Following initial validation studies using purified lectins, we profiled the targets of HMO probes in live mouse macrophages. Using this strategy, we mapped hundreds of HMO binding partners across multiple cellular compartments, including many known glycan-binding proteins as well as numerous proteins previously not known to bind glycans. We expect our findings to inform future investigations of the diverse roles of how HMOs may regulate protein function.
人乳寡糖（HMOs）是一类存在于人乳中的未结合可溶性聚糖，具有多种生物活性。尽管最近的研究揭示了HMOs的众多生物学功能（抗菌、抗炎和益生菌特性），但参与这些过程的受体和蛋白质结合伴侣尚未得到很好的表征。这在很大程度上可归因于可溶性聚糖 - 蛋白质相互作用的低亲和力和短暂性，使得无法使用传统表征技术来检测活细胞中的结合伴侣。在此，我们展示了使用合成的可光活化HMO探针，通过基于质谱的化学蛋白质组学在活细胞中捕获、富集和鉴定HMO蛋白质靶点。在使用纯化凝集素进行初步验证研究后，我们分析了活小鼠巨噬细胞中HMO探针的靶点。使用这种策略，我们在多个细胞区室中绘制了数百个HMO结合伴侣，包括许多已知的聚糖结合蛋白以及许多以前未知与聚糖结合的蛋白质。我们期望我们的发现为未来关于HMOs如何调节蛋白质功能的多种作用的研究提供信息。
