Genomic biology has motivated the search for new general techniques for detecting protein-protein interactions in living cells. We have developed a new class of techniques based on interaction-dependent probe ligation. The affinity between a probe ligase and its peptide substrate is tuned such that probe transfer from the ligase to the peptide occurs only when the proteins (A and B in the scheme below) to which they are fused bring them together. We have demonstrated the concept using both biotin ligase and lipoic acid ligase. A major advantage compared to protein complementation assays (such as BiFC based on fluorescent protein reconstitution) is that our reporter is non-trapping, such that A and B can dissociate after coming together. Furthermore, the ligase and peptide substrate have low intrinsic affinity, reducing the rate of false positives.
Imaging protein-protein interactions inside living cells via interaction-dependent fluorophore ligation.
Journal of the American Chemical Society 2011, 133, 19769-19776.
S. A. Slavoff, D. S. Liu, J. D. Cohen, and A. Y. Ting.
Protein-protein interaction detection in vitro and in cells by proximity biotinylation.
An engineered aryl azide ligase for site-specific mapping of protein-protein interactions through photocrosslinking.
Journal of the American Chemical Society 2008, 130, 9251-9253.
M. Fernandez-Suarez, T. S. Chen, and A. Y. Ting
Angewandte Chemie International Edition 2008, 47, 7018-7021.
H. Baruah, S. Puthenveetil, Y.-A. Choi, S. Shah, and A. Y. Ting.
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