ArticleNucleic acids research2010
Building blocks for protein interaction devices.
Article in Nucleic acids research, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed.
- Cohesin prevents local mixing of condensed euchromatic domains in living human cells.Nature genetics · 2026Article
- Cohesin prevents local mixing of condensed euchromatic domains in living human cells.bioRxiv : the preprint server for biology · 2026Article
- Orthogonal Synthetic Zippers as Protein Scaffolds.ACS omega · 2018Article
- Article
- Design and implementation of a biomolecular concentration tracker.ACS synthetic biology · 2015Article
- Detecting protein-protein interactions based on kinase-mediated growth induction of mammalian cells.Scientific reports · 2014Article
- Engineering of weak helper interactions for high-efficiency FRET probes.Nature methods · 2013Article
- Rapid titration of retroviral vectors using a β-lactamase protein fragment complementation assay.Gene therapy · 2013Article
- Directed surface attachment of nanomaterials via coiled-coil-driven self-assembly.Nanotechnology · 2012Article
- Engineering robust control of two-component system phosphotransfer using modular scaffolds.Proceedings of the National Academy of Sciences of the United States of America · 2012Article
- SYNZIP protein interaction toolbox: in vitro and in vivo specifications of heterospecific coiled-coil interaction domains.ACS synthetic biology · 2012Article
- A tricistronic human adrenodoxin reductase-adrenodoxin-cytochrome P450 27A1 vector system for substrate hydroxylation in Escherichia coli.Protein expression and purification · 2011Article
- Article
- A biobrick library for cloning custom eukaryotic plasmids.PloS one · 2011Article
- BioNetCAD: design, simulation and experimental validation of synthetic biochemical networks.Bioinformatics (Oxford, England) · 2010Article
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Authors and funding
5 authors.
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Abstract
Here, we propose a framework for the design of synthetic protein networks from modular protein-protein or protein-peptide interactions and provide a starter toolkit of protein building blocks. Our proof of concept experiments outline a general work flow for part-based protein systems engineering. We streamlined the iterative BioBrick cloning protocol and assembled 25 synthetic multidomain proteins each from seven standardized DNA fragments. A systematic screen revealed two main factors controlling protein expression in Escherichia coli: obstruction of translation initiation by mRNA secondary structure or toxicity of individual domains. Eventually, 13 proteins were purified for further characterization. Starting from well-established biotechnological tools, two general-purpose interaction input and two readout devices were built and characterized in vitro. Constitutive interaction input was achieved with a pair of synthetic leucine zippers. The second interaction was drug-controlled utilizing the rapamycin-induced binding of FRB(T2098L) to FKBP12. The interaction kinetics of both devices were analyzed by surface plasmon resonance. Readout was based on Förster resonance energy transfer between fluorescent proteins and was quantified for various combinations of input and output devices. Our results demonstrate the feasibility of parts-based protein synthetic biology. Additionally, we identify future challenges and limitations of modular design along with approaches to address them.
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