SYNZIP Protein Interaction Toolbox: in Vitro and in Vivo Specifications of Heterospecific Coiled-Coil Interaction Domains
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Keating_Synzip protein.pdf
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5.91 MB
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Author(s)
Thompson, Kenneth Evan
Bashor, Caleb
Lim, Wendell A.
Keating, Amy E.
Date Issued
February 2012
Journal
ACS Synthetic Biology
Publisher
American Chemical Society
Citation
Thompson, Kenneth Evan et al. “SYNZIP Protein Interaction Toolbox: In Vitro and in Vivo Specifications of Heterospecific Coiled-Coil Interaction Domains.” ACS Synthetic Biology 1.4 (2012): 118–129. Web. © 2012 American Chemical Society.
Version
Final published version
Abstract
The synthetic biology toolkit contains a growing number of parts for regulating transcription and translation, but very few that can be used to control protein association. Here we report characterization of 22 previously published heterospecific synthetic coiled-coil peptides called SYNZIPs. We present biophysical analysis of the oligomerization states, helix orientations, and affinities of 27 SYNZIP pairs. SYNZIP pairs were also tested for interaction in two cell-based assays. In a yeast two-hybrid screen, >85% of 253 comparable interactions were consistent with prior in vitro measurements made using coiled-coil microarrays. In a yeast-signaling assay controlled by coiled-coil mediated scaffolding, 12 SYNZIP pairs were successfully used to down-regulate the expression of a reporter gene following treatment with α-factor. Characterization of these interaction modules dramatically increases the number of available protein interaction parts for synthetic biology and should facilitate a wide range of molecular engineering applications. Summary characteristics of 27 SYNZIP peptide pairs are reported in specification sheets available in the Supporting Information and at the SYNZIP Web site [http://keatingweb.mit.edu/SYNZIP/].
MIT Department
Massachusetts Institute of Technology. Department of Biology
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DOI of Published Version
https://doi.org/10.1021/sb200015u
