SYNZIP Protein Interaction Toolbox: in Vitro and in Vivo Specifications of Heterospecific Coiled-Coil Interaction Domains
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 pre...
| Main Authors: | , , , |
|---|---|
| Other Authors: | |
| Format: | Article |
| Language: | en_US |
| Published: |
American Chemical Society
2012
|
| Online Access: | http://hdl.handle.net/1721.1/73587 https://orcid.org/0000-0003-4074-8980 |
| _version_ | 1811077786557743104 |
|---|---|
| author | Thompson, Kenneth Evan Bashor, Caleb Lim, Wendell A. Keating, Amy E. |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Thompson, Kenneth Evan Bashor, Caleb Lim, Wendell A. Keating, Amy E. |
| author_sort | Thompson, Kenneth Evan |
| collection | MIT |
| description | 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/]. |
| first_indexed | 2024-09-23T10:48:22Z |
| format | Article |
| id | mit-1721.1/73587 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T10:48:22Z |
| publishDate | 2012 |
| publisher | American Chemical Society |
| record_format | dspace |
| spelling | mit-1721.1/735872022-09-30T23:09:54Z SYNZIP Protein Interaction Toolbox: in Vitro and in Vivo Specifications of Heterospecific Coiled-Coil Interaction Domains Thompson, Kenneth Evan Bashor, Caleb Lim, Wendell A. Keating, Amy E. Massachusetts Institute of Technology. Department of Biology Thompson, Kenneth Evan Keating, Amy E. 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/]. National Science Foundation (U.S.) (NSF award MCB 0950233) National Institutes of Health (U.S.) (grant RO1 GM55040) National Institutes of Health (U.S.) (grant PN2 EY016546) National Institutes of Health (U.S.) (grant P50 GMO81879) National Science Foundation (U.S.). Synthetic Biology Engineering Research Center Howard Hughes Medical Institute 2012-10-04T13:32:07Z 2012-10-04T13:32:07Z 2012-02 2011-10 Article http://purl.org/eprint/type/JournalArticle 2161-5063 http://hdl.handle.net/1721.1/73587 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. https://orcid.org/0000-0003-4074-8980 en_US http://dx.doi.org/10.1021/sb200015u ACS Synthetic Biology Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf American Chemical Society PMC |
| spellingShingle | Thompson, Kenneth Evan Bashor, Caleb Lim, Wendell A. Keating, Amy E. 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 |
| title_full | SYNZIP Protein Interaction Toolbox: in Vitro and in Vivo Specifications of Heterospecific Coiled-Coil Interaction Domains |
| title_fullStr | SYNZIP Protein Interaction Toolbox: in Vitro and in Vivo Specifications of Heterospecific Coiled-Coil Interaction Domains |
| title_full_unstemmed | SYNZIP Protein Interaction Toolbox: in Vitro and in Vivo Specifications of Heterospecific Coiled-Coil Interaction Domains |
| title_short | SYNZIP Protein Interaction Toolbox: in Vitro and in Vivo Specifications of Heterospecific Coiled-Coil Interaction Domains |
| title_sort | synzip protein interaction toolbox in vitro and in vivo specifications of heterospecific coiled coil interaction domains |
| url | http://hdl.handle.net/1721.1/73587 https://orcid.org/0000-0003-4074-8980 |
| work_keys_str_mv | AT thompsonkennethevan synzipproteininteractiontoolboxinvitroandinvivospecificationsofheterospecificcoiledcoilinteractiondomains AT bashorcaleb synzipproteininteractiontoolboxinvitroandinvivospecificationsofheterospecificcoiledcoilinteractiondomains AT limwendella synzipproteininteractiontoolboxinvitroandinvivospecificationsofheterospecificcoiledcoilinteractiondomains AT keatingamye synzipproteininteractiontoolboxinvitroandinvivospecificationsofheterospecificcoiledcoilinteractiondomains |