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SYNZIP Protein Interaction Toolbox: in Vitro and in Vivo Specifications of Heterospecific Coiled-Coil Interaction Domains
[Image: see text] 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 call...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Chemical
Society
2012
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3339576/ https://www.ncbi.nlm.nih.gov/pubmed/22558529 http://dx.doi.org/10.1021/sb200015u |
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author | Thompson, Kenneth Evan Bashor, Caleb J. Lim, Wendell A. Keating, Amy E. |
author_facet | Thompson, Kenneth Evan Bashor, Caleb J. Lim, Wendell A. Keating, Amy E. |
author_sort | Thompson, Kenneth Evan |
collection | PubMed |
description | [Image: see text] 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/]. |
format | Online Article Text |
id | pubmed-3339576 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | American Chemical
Society |
record_format | MEDLINE/PubMed |
spelling | pubmed-33395762012-05-01 SYNZIP Protein Interaction Toolbox: in Vitro and in Vivo Specifications of Heterospecific Coiled-Coil Interaction Domains Thompson, Kenneth Evan Bashor, Caleb J. Lim, Wendell A. Keating, Amy E. ACS Synth Biol [Image: see text] 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/]. American Chemical Society 2012-02-02 2012-04-20 /pmc/articles/PMC3339576/ /pubmed/22558529 http://dx.doi.org/10.1021/sb200015u Text en Copyright © 2012 American Chemical Society http://pubs.acs.org This is an open-access article distributed under the ACS AuthorChoice Terms & Conditions. Any use of this article, must conform to the terms of that license which are available at http://pubs.acs.org. |
spellingShingle | Thompson, Kenneth Evan Bashor, Caleb J. 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 | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3339576/ https://www.ncbi.nlm.nih.gov/pubmed/22558529 http://dx.doi.org/10.1021/sb200015u |
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