ArticleNucleic acids research2007
High-precision mapping of protein protein interfaces: an integrated genetic strategy combining en masse mutagenesis and DNA-level parallel analysis on a yeast two-hybrid platform.
Article in Nucleic acids research, 2007. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
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Who cites it
9 citing papers in PubMed, 17 citations in OpenAlex.
- Generation of comprehensive transposon insertion mutant library for the model archaeon, Haloferax volcanii, and its use for gene discovery.BMC biology · 2014Article
- An assay to monitor the activity of DNA transposition complexes yields a general quality control measure for transpositional recombination reactions.Mobile genetic elements · 2014Article
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- Universal platform for quantitative analysis of DNA transposition.Mobile DNA · 2010Article
- Mso1p regulates membrane fusion through interactions with the putative N-peptide-binding area in Sec1p domain 1.Molecular biology of the cell · 2010Article
- A general method for scanning unnatural amino acid mutagenesis.ACS chemical biology · 2009Article
- Transposition-based method for the rapid generation of gene-targeting vectors to produce Cre/Flp-modifiable conditional knock-out mice.PloS one · 2009Article
- Bacteriophage Mu integration in yeast and mammalian genomes.Nucleic acids research · 2008Article
Corrections and comments
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Authors and funding
6 authors at 4 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Understanding networks of protein-protein interactions constitutes an essential component on a path towards comprehensive description of cell function. Whereas efficient techniques are readily available for the initial identification of interacting protein partners, practical strategies are lacking for the subsequent high-resolution mapping of regions involved in protein-protein interfaces. We present here a genetic strategy to accurately map interacting protein regions at amino acid precision. The system is based on parallel construction, sampling and analysis of a comprehensive insertion mutant library. The methodology integrates Mu in vitro transposition-based random pentapeptide mutagenesis of proteins, yeast two-hybrid screening and high-resolution genetic footprinting. The strategy is general and applicable to any interacting protein pair. We demonstrate the feasibility of the methodology by mapping the region in human JFC1 that interacts with Rab8A, and we show that the association is mediated by the Slp homology domain 1.
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