Protein–protein interactions monitored in mammalian cells via complementation of β-lactamase enzyme fragments
- 19 March 2002
- journal article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences of the United States of America
- Vol. 99 (6), 3469-3474
- https://doi.org/10.1073/pnas.062043699
Abstract
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.Keywords
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