Proteinâprotein interactions monitored in mammalian cells via complementation of β-lactamase enzyme fragments Export

@article{citeulike:966538, abstract = {10.1073/pnas.062043699 We have defined inactive \^{I}± and \"{I}‰ fragments of \^{I}²-lactamase that can complement to form a functional enzyme in both bacteria and mammalian cells, serving as a readout for the interaction of proteins fused to the fragments. Critical to this advance was the identification of a tripeptide, Asn-Gly-Arg, which when juxtaposed at the carboxyl terminus of the \^{I}± fragment increased complemented enzyme activity by up to 4 orders of magnitude. \^{I}²-Lactamase is well suited to monitoring constitutive and inducible protein interactions because it is small (29 kDa), monomeric, and assayable with a fluorescent cell-permeable substrate. The negligible background, the magnitude of induced signal caused by enzymatic amplification, and detection of signal within minutes are unparalleled in mammalian protein interaction detection systems published to date.}, author = {Wehrman, Tom and Kleaveland, Benjamin and Her, Jeng-Horng and Balint, Robert F. and Blau, Helen M.}, citeulike-article-id = {966538}, citeulike-linkout-0 = {http://dx.doi.org/10.1073/pnas.062043699}, citeulike-linkout-1 = {http://www.pnas.org/content/99/6/3469.abstract}, citeulike-linkout-2 = {http://www.pnas.org/content/99/6/3469.full.pdf}, citeulike-linkout-3 = {http://www.pnas.org/cgi/content/abstract/99/6/3469}, citeulike-linkout-4 = {http://view.ncbi.nlm.nih.gov/pubmed/11904411}, citeulike-linkout-5 = {http://www.hubmed.org/display.cgi?uids=11904411}, day = {19}, doi = {10.1073/pnas.062043699}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, keywords = {beta-lactamase, complementation, interaction, protein}, month = {March}, number = {6}, pages = {3469--3474}, posted-at = {2006-11-29 13:15:07}, priority = {2}, title = {Protein\^{a}€“protein interactions monitored in mammalian cells via complementation of \^{I}²-lactamase enzyme fragments}, url = {http://dx.doi.org/10.1073/pnas.062043699}, volume = {99}, year = {2002} }

TY - JOUR ID - citeulike:966538 L3 - citeulike-article-id:966538 N2 - 10.1073/pnas.062043699 We have defined inactive α and ω fragments of β-lactamase that can complement to form a functional enzyme in both bacteria and mammalian cells, serving as a readout for the interaction of proteins fused to the fragments. Critical to this advance was the identification of a tripeptide, Asn-Gly-Arg, which when juxtaposed at the carboxyl terminus of the α fragment increased complemented enzyme activity by up to 4 orders of magnitude. β-Lactamase is well suited to monitoring constitutive and inducible protein interactions because it is small (29 kDa), monomeric, and assayable with a fluorescent cell-permeable substrate. The negligible background, the magnitude of induced signal caused by enzymatic amplification, and detection of signal within minutes are unparalleled in mammalian protein interaction detection systems published to date. IS - 6 JF - Proceedings of the National Academy of Sciences of the United States of America EP - 3474 TI - Protein–protein interactions monitored in mammalian cells via complementation of β-lactamase enzyme fragments VL - 99 SP - 3469 KW - beta-lactamase KW - complementation KW - interaction KW - protein AU - Wehrman, Tom AU - Kleaveland, Benjamin AU - Her, Jeng-Horng AU - Balint, Robert AU - Blau, Helen PY - 2002/03/19/ UR - http://dx.doi.org/10.1073/pnas.062043699 ER -