Department of Genetic Medicine, Weill Cornell Medicine in Qatar, Doha, Qatar.
Cancer Med. 2022 Dec;11(24):4989-5000. doi: 10.1002/cam4.4805. Epub 2022 May 14.
Identify protein contact points between TP53 and minichromosome maintenance (MCM) complex proteins 2, 3, and 5 with high resolution allowing for potential novel Cancer drug design.
A next-generation sequencing-based protein-protein interaction method developed in our laboratory called AVA-Seq was applied to a gold-standard human protein interaction set. Proteins including TP53, MCM2, MCM3, MCM5, HSP90AA1, PCNA, NOD1, and others were sheared and ligated into the AVA-Seq system. Protein-protein interactions were then identified in both mild and stringent selective conditions.
Known interactions among MCM2, MCM3, and MCM5 were identified with the AVA-Seq system. The interacting regions detected between these three proteins overlap with the structural data of the MCM complex, and novel domains were identified with high resolution determined by multiple overlapping fragments. Fragments of wild type TP53 were shown to interact with MCM2, MCM3, and MCM5, and details on the location of the interactions were provided. Finally, a mini-network of known and novel cancer protein interactions was provided, which could have implications for fundamental changes in multiple cancers.
We provide a high-resolution mini-interactome that could direct novel drug targets and implicate possible effects of specific cancer mutations.
确定 TP53 与微小染色体维持（MCM）复合物蛋白 2、3 和 5 之间的蛋白质接触点，分辨率高，可用于潜在的新型癌症药物设计。
应用我们实验室开发的下一代测序蛋白-蛋白相互作用方法 AVA-Seq 对黄金标准的人类蛋白相互作用组进行了分析。包括 TP53、MCM2、MCM3、MCM5、HSP90AA1、PCNA、NOD1 等在内的蛋白质被剪切并连接到 AVA-Seq 系统中。然后在温和和严格的选择性条件下鉴定蛋白-蛋白相互作用。
在 AVA-Seq 系统中鉴定到了 MCM2、MCM3 和 MCM5 之间的已知相互作用。检测到的这三个蛋白之间的相互作用区域与 MCM 复合物的结构数据重叠，并通过多个重叠片段以高分辨率鉴定出了新的结构域。野生型 TP53 的片段被证明与 MCM2、MCM3 和 MCM5 相互作用，并提供了相互作用位置的详细信息。最后，提供了一个已知和新型癌症蛋白相互作用的迷你网络，这可能对多种癌症的根本变化产生影响。
我们提供了一个高分辨率的迷你互作组，可直接作为新型药物靶点，并暗示特定癌症突变可能产生的影响。
