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Book of Abstracts: Albany 2007

category image Albany 2007
Conversation 15
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
  1. Burz, D. S., Dutta, K., Cowburn, D., Shekhtman, A. Nature Protocols 1, 146-152 (2006).
  2. Burz, D. S., Dutta, K., Cowburn, D., Shekhtman, A. Nature Methods 3, 91-93 (2006).

David. S. Burz
Alexander Shekhtman

State University of New York at Albany
Department of Chemistry
1400 Washington Ave
Albany, NY 12222

Email: as979494@albany.edu