Protein fragment complementation assays

Detecting interactions by protein fragment complementation assays... 

Protein fragment complementation assays are based on an enzyme reassembly strategy whereby a protein-protein interaction promotes the efficient refolding and complementation of enzyme fragments to restore an active enzyme. The approach was initially developed using the reconstitution of ubiquitin as a sensor for protein-protein interactions (Johnsson and Varshavsky, 1994). Ubiquitin is a 76 amino acid protein that... 

Remy, I., and Michnick, S. W. (1999). Clonal selection and in vivo quantitation of protein interactions with protein-fragment complementation assays. Proc. Natl. Acad. Sci. USA 96, 5394-5399. 

Systematic Analysis of Complex Signal Transduction Pathways Using Protein Fragment Complementation Assays... 

Michnick SW, Ear PH, Manderson EN et al (2007) Universal strategies in research and drug discovery based on protein-fragment complementation assays. Nat Rev Drug Discov 6 569-582... 

Examples of such protein-protein interaction selection systems are phage display (Smith, 1985 Winter et al., 1994), display on other viruses (Kasahara et al., 1994), bacterial surface display (Georgiou et al., 1993 Daugherty et al., 1999), yeast display (Kieke et al., 1997 Boder and Wittrup, 1997), the yeast two hybrid system (Fields and Song, 1989 Chein et al., 1991), and protein-fragment complementation assays (Pelletier et al., 1998). These methods all contain a necessary in vivo step, which has a number of disadvantages that will be discussed in the following sections. 

Yu H, West M, Keon BH, Bilter GK, Owens S, Lamerdin J, Westwick JK. Measuring drug action in the cellular context using protein-fragment complementation assays. Assay Drug Dev. Technol. 2003, 1, 811-822. 

De A, Gambhir SS. Noninvasive imaging of protein-protein interactions from live cells and living snbjects nsing biolnminescence resonance energy transfer. Faseb J 2005 19 2017-2019. Michnick SW, Ear PH, Manderson EN, Remy I, Stefan E. Universal strategies in research and dmg discovery based on protein-fragment complementation assays. Nat. Rev. Dmg Dis-cov. 2007 6 569-582. 

Remy 1, Campbell-Valois FX, Michnick SW. Detection of protein-protein interactions using a simple survival protein-fragment complementation assay based on the enzyme dihydrofolate reductase. Nat. Protoc. 2007 2 2120-2125. 

Campbell-Valois FX, Michnick SW. Synthesis of degenerated libraries of the ras-binding domain of raf and rapid selection of fast-folding and stable clones with the dihydrofolate reductase protein fragment complementation assay. Methods Mol. Biol. 2007 352 249-274. 

Mossner, E., Koch, H., Pliickthun, A. Fast selection of antibodies without antigen purification adaption of the protein fragment complementation assay to select antigen-antibody pairs./. Mol. Biol. 2001, 308, 115-122. 

I. Remy, S.W. Michnick, Mapping biochemical networks with protein-fragment complementation assays, Methods Mol. Biol. 2004, 261, 411-426. 

Galameau A, Primeau M, Trudeau L-E, Michnick S. -Lactamase protein fragment complementation assays as in vivo and in vitro sensors of protein-protein interactions. Nat Biotechnol 2002 20 619-622. 

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