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. Export

@article{citeulike:1577081, 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.}, address = {Program in Cellular Biotechnology, Institute of Biotechnology, Viikki Biocenter, University of Helsinki, Institute of Medical Technology, University of Tampere, Research Unit, Tampere University Hospital, Tampere and Division of Genetics and Physiology, Department of Biology, University of Turku, Finland.}, author = {Pajunen, Maria and Turakainen, Hilkka and Poussu, Eini and Per\"{a}nen, Johan and Vihinen, Mauno and Savilahti, Harri}, citeulike-article-id = {1577081}, citeulike-linkout-0 = {http://dx.doi.org/10.1093/nar/gkm563}, citeulike-linkout-1 = {http://nar.oxfordjournals.org/cgi/content/abstract/35/16/e103}, citeulike-linkout-2 = {http://view.ncbi.nlm.nih.gov/pubmed/17702760}, citeulike-linkout-3 = {http://www.hubmed.org/display.cgi?uids=17702760}, day = {15}, doi = {10.1093/nar/gkm563}, issn = {1362-4962}, journal = {Nucleic Acids Res}, keywords = {interaction, mapping, mutatgenesis, protein, two-hybrid, yeast}, month = {August}, posted-at = {2007-08-20 17:22:32}, priority = {3}, title = {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.}, url = {http://dx.doi.org/10.1093/nar/gkm563}, year = {2007} }

TY - JOUR ID - citeulike:1577081 L3 - citeulike-article-id:1577081 N2 - 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. JF - Nucleic Acids Res SN - 1362-4962 AD - Program in Cellular Biotechnology, Institute of Biotechnology, Viikki Biocenter, University of Helsinki, Institute of Medical Technology, University of Tampere, Research Unit, Tampere University Hospital, Tampere and Division of Genetics and Physiology, Department of Biology, University of Turku, Finland. TI - 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. KW - interaction KW - mapping KW - mutatgenesis KW - protein KW - two-hybrid KW - yeast AU - Pajunen, Maria AU - Turakainen, Hilkka AU - Poussu, Eini AU - Peränen, Johan AU - Vihinen, Mauno AU - Savilahti, Harri PY - 2007/08/15/ UR - http://dx.doi.org/10.1093/nar/gkm563 ER -