美国斯克利普斯研究所Cravatt, Benjamin F.团队报道了癌症细胞中胰蛋白酶-丙烯酰胺-蛋白质相互作用的多重化学蛋白质组学图谱。相关研究成果于2024年8月13日发表在《自然—化学》。

共价化学是一种扩展人类蛋白质组可连接性的多功能方法。基于活性的蛋白质谱（ABPP）可以通过与广泛反应的探针竞争来推断亲电化合物修饰的特定残基。然而，这种残基导向平台在多大程度上完全评估了细胞中亲电化合物的蛋白质靶标尚不清楚。

该文中，研究人员评估了一种互补的蛋白质导向ABPP方法，该方法鉴定了与炔基化手性亲电化合物（称为立体探针）具有立体选择性反应的蛋白质。整合来自用胰蛋白酶-丙烯酰胺立体探针处理的癌症细胞的蛋白质和半胱氨酸导向数据，揭示了总体上相关性良好的连接性图谱和突出的特征，如蛋白质大小和含半胱氨酸肽的蛋白型，这些特征解释了每个ABPP平台中的差距。

总的来说，研究人员确定了>300种结构和功能不同的蛋白质的立体探针结合事件，包括立体选择性和位点特异性破坏MAD2L1BP与纺锤体组装检查点复合物相互作用的导致癌症细胞有丝分裂退出延迟的化合物。

Title: Multi-tiered chemical proteomic maps of tryptoline acrylamide–protein interactions in cancer cells

Author: Njomen, Evert, Hayward, Rachel E., DeMeester, Kristen E., Ogasawara, Daisuke, Dix, Melissa M., Nguyen, Tracey, Ashby, Paige, Simon, Gabriel M., Schreiber, Stuart L., Melillo, Bruno, Cravatt, Benjamin F.

Issue&Volume: 2024-08-13

Abstract: Covalent chemistry is a versatile approach for expanding the ligandability of the human proteome. Activity-based protein profiling (ABPP) can infer the specific residues modified by electrophilic compounds through competition with broadly reactive probes. However, the extent to which such residue-directed platforms fully assess the protein targets of electrophilic compounds in cells remains unclear. Here we evaluate a complementary protein-directed ABPP method that identifies proteins showing stereoselective reactivity with alkynylated, chiral electrophilic compounds—termed stereoprobes. Integration of protein- and cysteine-directed data from cancer cells treated with tryptoline acrylamide stereoprobes revealed generally well-correlated ligandability maps and highlighted features, such as protein size and the proteotypicity of cysteine-containing peptides, that explain gaps in each ABPP platform. In total, we identified stereoprobe binding events for >300 structurally and functionally diverse proteins, including compounds that stereoselectively and site-specifically disrupt MAD2L1BP interactions with the spindle assembly checkpoint complex leading to delayed mitotic exit in cancer cells.

DOI: 10.1038/s41557-024-01601-1

Source: https://www.nature.com/articles/s41557-024-01601-1