Protein-protein interactions monitored in mammalian cells via complementation of beta -lactamase enzyme fragments

Proceedings of the National Academy of Sciences of the United States of America
Tom S WehrmanHelen M Blau

Abstract

We have defined inactive alpha and omega fragments of beta-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 alpha fragment increased complemented enzyme activity by up to 4 orders of magnitude. beta-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.

References

Feb 21, 1991·Nature·S R AdamsR Y Tsien
Sep 1, 1994·Immunology Today·F H DurieR J Noelle
May 23, 1995·Proceedings of the National Academy of Sciences of the United States of America·J ChenS L Schreiber
Jul 18, 1995·Proceedings of the National Academy of Sciences of the United States of America·I RivièreR C Mulligan
May 14, 1996·Proceedings of the National Academy of Sciences of the United States of America·P J BelshawS L Schreiber
Apr 1, 1997·Current Opinion in Cell Biology·M KarinE Zandi
Aug 5, 1997·Proceedings of the National Academy of Sciences of the United States of America·F RossiH M Blau
Jun 6, 1998·Cellular and Molecular Life Sciences : CMLS·A PhilipponJ M Frère
Feb 4, 1999·Biochemistry·S McManus-Munoz, M W Crowder
May 13, 1999·Proceedings of the National Academy of Sciences of the United States of America·I Remy, S W Michnick
Feb 5, 2000·Nature Biotechnology·B T BlakelyH M Blau

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Citations

Aug 31, 2002·Angewandte Chemie·Eric A Althoff, Virginia W Cornish
Dec 3, 2008·Molecular Imaging and Biology : MIB : the Official Publication of the Academy of Molecular Imaging·C T ChanS S Gambhir
Dec 10, 2003·Current Opinion in Biotechnology·Stephen W Michnick
Feb 5, 2003·Current Opinion in Biotechnology·Daniel Figeys
Mar 9, 2004·Drug Discovery Today·Stephen W Michnick
Oct 16, 2003·Current Opinion in Pharmacology·Michael Entzeroth
Nov 11, 2009·Molecular Pharmaceutics·Ann-Marie BroomeJames P Basilion
Dec 8, 2004·Nature Biotechnology·Stéphanie CabantousGeoffrey S Waldo
Oct 31, 2006·Nature Methods·Michael C WehrMoritz J Rossner
Jun 30, 2007·Nature Reviews. Drug Discovery·Stephen W MichnickEduard Stefan
Feb 24, 2009·Proceedings of the National Academy of Sciences of the United States of America·Dujduan Waraho, Matthew P DeLisa
Apr 20, 2004·Assay and Drug Development Technologies·Richard M Eglen
May 6, 2008·Protein Engineering, Design & Selection : PEDS·M VandevenneP Filée
Oct 3, 2008·Protein Engineering, Design & Selection : PEDS·Paola SeccoDaniele Sblattero
Dec 12, 2012·Protein Engineering, Design & Selection : PEDS·Janina SpeckKristian M Müller
Apr 28, 2007·Annual Review of Biomedical Engineering·Victor VillalobosDavid Piwnica-Worms
Jan 1, 2008·Biotechnology & Genetic Engineering Reviews·Tingting JiangJianghong Rao
Dec 3, 2003·Proceedings of the National Academy of Sciences of the United States of America·Sumitaka HasegawaJianghong Rao
Nov 5, 2002·Proceedings of the National Academy of Sciences of the United States of America·James M SpottsMichael E Greenberg
Oct 19, 2007·Proceedings of the National Academy of Sciences of the United States of America·E StefanS W Michnick
Dec 7, 2006·Proceedings of the National Academy of Sciences of the United States of America·T S WehrmanH M Blau
Aug 1, 2007·Proceedings of the National Academy of Sciences of the United States of America·Youngsoo Jun, William Wickner
Sep 21, 2007·Proceedings of the National Academy of Sciences of the United States of America·Xuehui ChenTimothy A Springer
Nov 20, 2002·Proceedings of the National Academy of Sciences of the United States of America·R PaulmuruganS S Gambhir
Aug 1, 2012·Stem Cell Reviews and Reports·Maram YousefiGhasem Hosseini Salekdeh
Mar 12, 2008·The Journal of Cell Biology·Kostandin V PajciniHelen M Blau
Dec 14, 2005·Journal of Pharmacological and Toxicological Methods·Charlotte Harrison, Piet H van der Graaf
Nov 13, 2007·Biophysical Journal·Lydia M Contreras MartínezMatthew P DeLisa
Oct 3, 2003·Cancer Cell·Mary Ellen MartinJay L Hess
Mar 25, 2008·Biochemical and Biophysical Research Communications·Rachel de las HerasChang-Yi Huang
Apr 25, 2006·Biochemical and Biophysical Research Communications·Chenjie XuJianghong Rao
Jan 24, 2009·Protein Science : a Publication of the Protein Society·Yong ChenZhanglin Lin
Dec 23, 2015·Protein Science : a Publication of the Protein Society·Tsz-Leung ToXiaokun Shu

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