SURF 2013: Designing phosphorylation sensitive protein domains for use in synthetic circuits

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2013 SURF project description

  • Mentor: Richard Murray
  • Co-mentor: Emzo de los Santos

Synthetic biology aims take advantage of biological process to engineer systems and devices with novel and useful functions. Since it is still in its infancy, much work is necessary in order to build a framework to make it easier to design using biological parts. This project aims to contribute to this framework and expand the synthetic biology toolkit.

The project involves characterizing peptide domains that are designed to have thermostability differences in the phgosphorylated and non-phosphorylated states. By inserting these domains into different proteins, we believe that we can get a general method for stimulus-dependent modulation of protein activity. This work builds on previous work that has previously shown modulation in activity of enzymes through insertion of light-sensitive [1] and chemically-sensitive domains [2]. Phosphorylation events are widely used in natural systems to control the dynamics of gene networks. Circuits including proteins that using phosphorylation as a trigger for modulation of activity should be able to respond on a faster time scale than traditional synthetic circuits.

Biochemical characterization will be done both of the domains on their own and inserted into at least one enzyme. Mathematical modeling and characterization of circuits that use this mechanism can also be performed.

References

  1. Reynolds, Kimberly A, Richard N McLaughlin, and Rama Ranganathan. “Hot Spots for Allosteric Regulation on Protein Surfaces..” Cell 147.7 (2011): 1564–1575.
  2. Erster, O, M Eisenstein, and M Liscovitch. “Ligand Interaction Scan: a General Method for Engineering Ligand- Sensitive Protein Alleles.” Nature methods (2007)