Paulmurugan R, Umezawa Y, Gambhir S S
Crump Institute for Molecular Imaging, Department of Molecular and Medical Pharmacology, University of California Los Angeles-Jonsson Comprehensive Cancer Center, 90095-1770, USA.
Proc Natl Acad Sci U S A. 2002 Nov 26;99(24):15608-13. doi: 10.1073/pnas.242594299. Epub 2002 Nov 18.
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.
在本研究中，我们开发了生物发光成像策略，通过使用冷却电荷耦合器件相机和分裂报告技术，在活体小鼠中对蛋白质 - 蛋白质相互作用进行非侵入性定量成像。我们验证了由两个强相互作用蛋白MyoD和Id的相互作用驱动的分裂萤火虫荧光素酶蛋白的互补和内含肽介导的重组。我们使用细胞的瞬时转染，并在细胞培养物以及植入活体小鼠的细胞中诱导基因表达后，对MyoD - Id相互作用进行成像。研究活体受试者中蛋白质 - 蛋白质相互作用的技术将有助于研究细胞网络，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。
