Intracellular protein interaction mapping with FRET hybrids.

@article{citeulike:1151552, abstract = {A quantitative methodology was developed to identify protein interactions in a broad range of cell types by using FRET between fluorescent proteins. Genetic fusions of a target receptor to a FRET acceptor and a large library of candidate peptide ligands to a FRET donor enabled high-throughput optical screening for optimal interaction partners in the cytoplasm of Escherichia coli. Flow cytometric screening identified a panel of peptide ligands capable of recognizing the target receptors in the intracellular environment. For both SH3 and PDZ domain-type target receptors, physiologically meaningful consensus sequences were apparent among the isolated ligands. The relative dissociation constants of interacting partners could be measured directly by using a dilution series of cell lysates containing FRET hybrids, providing a previously undescribed high-throughput approach to rank the affinity of many interaction partners. FRET hybrid interaction screening provides a powerful tool to discover protein ligands in the cellular context with potential applications to a wide variety of eukaryotic cell types.}, address = {Department of Chemical Engineering, University of California-Santa Barbara, Santa Barbara, CA 93106, USA.}, author = {You, X. and Nguyen, A. W. and Jabaiah, A. and Sheff, M. A. and Thorn, K. S. and Daugherty, P. S. }, citeulike-article-id = {1151552}, doi = {http://dx.doi.org/10.1073/pnas.0605422103}, issn = {0027-8424}, journal = {Proc Natl Acad Sci U S A}, month = {December}, number = {49}, pages = {18458--18463}, posted-at = {2008-04-26 10:34:49}, priority = {2}, title = {Intracellular protein interaction mapping with FRET hybrids.}, url = {http://dx.doi.org/10.1073/pnas.0605422103}, volume = {103}, year = {2006} }

TY - JOUR ID - citeulike:1151552 L3 - citeulike-article-id:1151552 TI - Intracellular protein interaction mapping with FRET hybrids. JF - Proc Natl Acad Sci U S A VL - 103 IS - 49 SP - 18458 EP - 18463 AD - Department of Chemical Engineering, University of California-Santa Barbara, Santa Barbara, CA 93106, USA. SN - 0027-8424 N2 - A quantitative methodology was developed to identify protein interactions in a broad range of cell types by using FRET between fluorescent proteins. Genetic fusions of a target receptor to a FRET acceptor and a large library of candidate peptide ligands to a FRET donor enabled high-throughput optical screening for optimal interaction partners in the cytoplasm of Escherichia coli. Flow cytometric screening identified a panel of peptide ligands capable of recognizing the target receptors in the intracellular environment. For both SH3 and PDZ domain-type target receptors, physiologically meaningful consensus sequences were apparent among the isolated ligands. The relative dissociation constants of interacting partners could be measured directly by using a dilution series of cell lysates containing FRET hybrids, providing a previously undescribed high-throughput approach to rank the affinity of many interaction partners. FRET hybrid interaction screening provides a powerful tool to discover protein ligands in the cellular context with potential applications to a wide variety of eukaryotic cell types. AU - You, X AU - Nguyen, AW AU - Jabaiah, A AU - Sheff, MA AU - Thorn, KS AU - Daugherty, PS PY - 2006/12/05/ UR - http://dx.doi.org/10.1073/pnas.0605422103 ER -