Control of protein signaling using a computationally designed GTPase/GEF orthogonal pair

Gregory T. Kapp, Sen Liu, Amelie Stein, Derek T. Wong, Attila Remeńyi, Brian J. Yeh, James S. Fraser, Jack Taunton, Wendell A. Lim, Tanja Kortemme

Research output: Contribution to journalArticle

59 Citations (Scopus)


Signaling pathways depend on regulatory protein-protein interactions; controlling these interactions in cells has important applications for reengineering biological functions. As many regulatory proteins are modular, considerable progress in engineering signaling circuits has been made by recombining commonly occurring domains. Our ability to predictably engineer cellular functions, however, is constrained by complex crosstalk observed in naturally occurring domains. Here we demonstrate a strategy for improving and simplifying protein network engineering: using computational design to create orthogonal (non-crossreacting) protein-protein interfaces. We validated the design of the interface between a key signaling protein, the GTPase Cdc42, and its activator, Intersectin, biochemically and by solving the crystal structure of the engineered complex. The designed GTPase (orthoCdc42) is activated exclusively by its engineered cognate partner (orthoIntersectin), but maintains the ability to interface with other GTPase signaling circuit components in vitro. In mammalian cells, orthoCdc42 activity can be regulated by orthoIntersectin, but not wild-type Intersectin, showing that the designed interaction can trigger complex processes. Computational design of protein interfaces thus promises to provide specific components that facilitate the predictable engineering of cellular functions.

Original languageEnglish
Pages (from-to)5277-5282
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Issue number14
Publication statusPublished - Apr 3 2012



  • Computational modeling and design
  • Signal transduction
  • Synthetic biology

ASJC Scopus subject areas

  • General

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