title: An infrared reporter to detect spatiotemporal dynamics of protein-protein interactions.
abstract: We report a protein-fragment complementation assay (PCA) based on the engineered Deinococcus radiodurans infrared fluorescent protein IFP1.4. Unlike previous fluorescent protein PCAs, the IFP PCA is reversible, allowing analysis of spatiotemporal dynamics of hormone-induced signaling complexes in living yeast and mammalian cells at nanometer resolution. The inherently low background of infrared fluorescence permitted detection of subcellular reorganization of a signaling complex expressed at low abundance.
authors.affiliation: Département de Biochimie, Université de Montréal, Montréal, Québec, Canada.
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Tchekanda
authors.affiliation: Département de Biochimie, Université de Montréal, Montréal, Québec, Canada.
authors.forename: Durga
authors.initials: D
authors.lastname: Sivanesan
authors.affiliation: Département de Biochimie, Université de Montréal, Montréal, Québec, Canada.
authors.forename: Stephen W
authors.initials: SW
authors.lastname: Michnick
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D002478:Cells, Cultured; D034301:Deinococcus / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D007259:Infrared Rays; D008958:Models, Molecular; D062211:Spatio-Temporal Analysis
chemical_list: D001426:Bacterial Proteins; D005456:Fluorescent Dyes
references.citation: Proc Natl Acad Sci U S A. 2007 Oct 23;104(43):16916-21
references.pmid: 17942691
references.citation: Nat Cell Biol. 2000 Jan;2(1):25-9
references.pmid: 10620803
references.citation: Chem Biol. 2013 Aug 22;20(8):1078-86
references.pmid: 23891149
references.citation: Nat Methods. 2009 Nov;6(11):813-6
references.pmid: 19820714
references.citation: PLoS One. 2010 Feb 16;5(2):e9225
references.pmid: 20169075
references.citation: J Biol Chem. 2012 Mar 2;287(10):7000-9
references.pmid: 22210774
references.citation: Methods Enzymol. 2000;328:208-30
references.pmid: 11075347
references.citation: EMBO J. 1994 Sep 15;13(18):4269-77
references.pmid: 7925272
references.citation: Nature. 2013 Jul 11;499(7457):166-71
references.pmid: 23846654
references.citation: Mol Cell. 2002 Apr;9(4):789-98
references.pmid: 11983170
references.citation: Proc Natl Acad Sci U S A. 1994 Oct 25;91(22):10340-4
references.pmid: 7937952
references.citation: Nat Rev Mol Cell Biol. 2002 Sep;3(9):639-50
references.pmid: 12209124
references.citation: Nat Methods. 2006 Dec;3(12):977-9
references.pmid: 17099704
references.citation: Science. 2009 May 8;324(5928):804-7
references.pmid: 19423828
references.citation: Biochemistry. 2007 Aug 14;46(32):9283-91
references.pmid: 17658893
references.citation: Science. 2008 Jun 13;320(5882):1465-70
references.pmid: 18467557
references.citation: Cell. 1997 Mar 7;88(5):593-602
references.pmid: 9054499
references.citation: Nature. 2010 May 6;465(7294):101-5
references.pmid: 20400943
references.citation: Gene. 1995 Apr 14;156(1):119-22
references.pmid: 7737504
references.citation: BMC Biotechnol. 2012 Nov 23;12:89
references.pmid: 23176158
references.citation: Nat Struct Biol. 1996 Mar;3(3):244-51
references.pmid: 8605626
references.citation: Biophys J. 2009 Feb 18;96(4):1508-14
references.pmid: 19217867
references.citation: Nature. 2005 Nov 17;438(7066):325-31
references.pmid: 16292304
references.citation: Yeast. 1999 Oct;15(14):1541-53
references.pmid: 10514571
references.citation: Biochim Biophys Acta. 2004 Mar 11;1697(1-2):259-69
references.pmid: 15023366
references.citation: Nat Chem Biol. 2006 Jun;2(6):329-37
references.pmid: 16680159
references.citation: Nat Rev Drug Discov. 2007 Jul;6(7):569-82
references.pmid: 17599086