Noninvasive imaging of protein-protein interactions in living subjects by using reporter protein complementation and reconstitution strategies
Noninvasive imaging of protein-protein interactions in living subjects by using reporter protein complementation and reconstitution strategies
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DOI：
10.1073/pnas.242594299
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发表时间：
2002-11-26
影响因子：
11.1
通讯作者：
Gambhir, SS
中科院分区：
文献类型：
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作者：
Paulmurugan, R;Umezawa, Y;Gambhir, SS
在这项研究中，我们已经开发了生物发光成像策略，非侵入性和定量成像蛋白质-蛋白质相互作用在活的小鼠通过使用冷却的电荷耦合器件相机和分裂报告技术。我们验证两个互补和内含肽介导的重组分裂萤火虫荧光素酶蛋白驱动的两个强烈相互作用的蛋白质，MyoD和Id的相互作用。我们使用瞬时转染细胞和图像MyoD-Id的相互作用后，在细胞培养和细胞植入到活的小鼠中的基因表达诱导。研究活体中蛋白质-蛋白质相互作用的技术将允许研究细胞网络，包括信号转导途径，以及用于调节蛋白质-蛋白质相互作用的药物的开发和优化。
In this study we have developed bioluminescence-imaging strategies to noninvasively and quantitatively image protein-protein interactions in living mice by using a cooled charge-coupled device camera and split reporter technology. We validate both complementation and intein-mediated reconstitution of split firefly luciferase proteins driven by the interaction of two strongly interacting proteins, MyoD and Id. We use transient transfection of cells and image MyoD-Id interaction after induction of gene expression in cell culture and in cells implanted into living mice. Techniques to study protein-protein interactions in living subjects will allow the study of cellular networks, including signal transduction pathways, as well as development and optimization of pharmaceuticals for modulating protein-protein interactions.