期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：2002
卷号：99
期号：5
页码：3105-3110
DOI：10.1073/pnas.052710999
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：We are developing methods to image molecular and cellular events in living subjects. In this study, we validate imaging of protein--protein interactions in living mice by using bioluminescent optical imaging. We use the well studied yeast two-hybrid system adapted for mammalian cells and modify it to be inducible. We employ the NF-{kappa}B promoter to drive expression of two fusion proteins (VP16-MyoD and GAL4-ID). We modulate the NF-{kappa}B promoter through tumor necrosis factor . Firefly luciferase reporter gene expression is driven by the interaction of MyoD and ID through a transcriptional activation strategy. We demonstrate the ability to detect this induced protein-protein interaction in cell culture and image this induced interaction in living mice by using transiently transfected cells. The current approach will be a valuable and potentially generalizable tool to noninvasively and quantitatively image protein-protein interactions in living subjects. The approaches validated should have important implications for the study of protein-protein interactions in cells maintained in their natural in vivo environment as well as for the in vivo evaluation of new pharmaceuticals targeted to modulate protein-protein interactions.
