Book of Abstracts: Albany 2007
June 19-23 2007
New Method of Mapping STructural INTeractions using in-cell NMR spectroscopy (STINT-NMR)
We describe a high-throughput in-cell NMR-based method for mapping the STructural changes that accompany protein-protein INTeractions in vivo (STINT-NMR). The method entails sequentially expressing two (or more) proteins within a single bacterial cell in a time-controlled manner and monitoring their interactions using in-cell NMR spectroscopy. The resulting NMR spectra provide a complete titration of the interaction and define structural details of the interacting surfaces at atomic resolution. Unlike the case where interacting proteins are simultaneously over-expressed, the NMR spectral complexity is minimized since only the target protein is labeled with NMR active nuclei leaving the interactor protein(s) cryptic. The method provides a structural foundation for proteomic studies and can be combined with genetic and molecular screens for high-throughput selection.
References and Footnotes
David. S. Burz
State University of New York at Albany