# SYNZIP Protein Interaction Toolbox: in Vitro and in Vivo Specifications of Heterospecific Coiled-Coil Interaction Domains.

 Title SYNZIP Protein Interaction Toolbox: in Vitro and in Vivo Specifications of Heterospecific Coiled-Coil Interaction Domains. Publication Type Journal Article Year of Publication 2012 Authors Thompson KEvan, Bashor CJ, Lim Wa, Keating AE Journal ACS synthetic biology Volume 1 Pagination 118–129 Date Published apr ISSN 2161-5063 Keywords behaviors and functions, coiled coil, engineer living, he central goal in, heterodimer, most common approach is, protein, protein interaction, set of standard, synthetic biology, synthetic biology is to, systems that exhibit new, the, to start with a 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 $\alpha$-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/]. URL http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=3339576&tool=pmcentrez&rendertype=abstract DOI 10.1021/sb200015u