Cell-free cotranslation and selection using in vitro virus for high-throughput analysis of protein-protein interactions and complexes

@article{citeulike:2988184, abstract = {We have developed a simple and totally in vitro selection procedure based on cell-free cotranslation using a highly stable and efficient in vitro virus (IVV). Cell-free cotranslation of tagged bait and prey proteins is advantageous for the formation of protein complexes and allows high-throughput analysis of protein-protein interactions (PPI) as a result of providing in vitro instead of in vivo preparation of bait proteins. The use of plural selection rounds and a two-step purification of the IVV selection, followed by in vitro post-selection, is advantageous for decreasing false positives. In a single experiment using bait Fos, more than 10 interactors, including not only direct, but also indirect interactions, were enriched. Further, previously unidentified proteins containing novel leucine zipper (L-ZIP) motifs with minimal binding sites identified by sequence alignment as functional elements were detected as a result of using a randomly primed cDNA library. Thus, we consider that this simple IVV selection system based on cell-free cotranslation could be applicable to high-throughput and comprehensive analysis of PPI and complexes in large-scale settings involving parallel bait proteins. 10.1101/gr.3510505}, author = {Miyamoto-Sato, Etsuko and Ishizaka, Masamichi and Horisawa, Kenichi and Tateyama, Seiji and Takashima, Hideaki and Fuse, Shinichiro and Sue, Kaori and Hirai, Naoya and Masuoka, Kazuyo and Yanagawa, Hiroshi }, citeulike-article-id = {2988184}, doi = {http://dx.doi.org/10.1101/gr.3510505}, journal = {Genome Res.}, keywords = {termpaper}, month = {May}, number = {5}, pages = {710--717}, posted-at = {2008-07-11 13:58:20}, priority = {2}, title = {Cell-free cotranslation and selection using in vitro virus for high-throughput analysis of protein-protein interactions and complexes}, url = {http://dx.doi.org/10.1101/gr.3510505}, volume = {15}, year = {2005} }

TY - JOUR ID - citeulike:2988184 L3 - citeulike-article-id:2988184 TI - Cell-free cotranslation and selection using in vitro virus for high-throughput analysis of protein-protein interactions and complexes JF - Genome Res. VL - 15 IS - 5 SP - 710 EP - 717 N2 - We have developed a simple and totally in vitro selection procedure based on cell-free cotranslation using a highly stable and efficient in vitro virus (IVV). Cell-free cotranslation of tagged bait and prey proteins is advantageous for the formation of protein complexes and allows high-throughput analysis of protein-protein interactions (PPI) as a result of providing in vitro instead of in vivo preparation of bait proteins. The use of plural selection rounds and a two-step purification of the IVV selection, followed by in vitro post-selection, is advantageous for decreasing false positives. In a single experiment using bait Fos, more than 10 interactors, including not only direct, but also indirect interactions, were enriched. Further, previously unidentified proteins containing novel leucine zipper (L-ZIP) motifs with minimal binding sites identified by sequence alignment as functional elements were detected as a result of using a randomly primed cDNA library. Thus, we consider that this simple IVV selection system based on cell-free cotranslation could be applicable to high-throughput and comprehensive analysis of PPI and complexes in large-scale settings involving parallel bait proteins. 10.1101/gr.3510505 KW - termpaper AU - Miyamoto-Sato, Etsuko AU - Ishizaka, Masamichi AU - Horisawa, Kenichi AU - Tateyama, Seiji AU - Takashima, Hideaki AU - Fuse, Shinichiro AU - Sue, Kaori AU - Hirai, Naoya AU - Masuoka, Kazuyo AU - Yanagawa, Hiroshi PY - 2005/05/01/ UR - http://dx.doi.org/10.1101/gr.3510505 ER -