title: A sensor for quantification of macromolecular crowding in living cells.
abstract: Macromolecular crowding in cells influences processes such as folding, association and diffusion of proteins and polynucleic acids. Direct spatiotemporal readout of crowding would be a powerful approach for unraveling the structure of the cytoplasm and determining the impact of excluded volume on protein function in living cells. Here, we introduce a genetically encodable fluorescence resonance energy transfer (FRET) sensor for quantifying macromolecular crowding and discuss our application of the sensor in bacterial and mammalian cells.
authors: 1] Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Groningen, the Netherlands. [2] Zernike Institute for Advanced Materials, University of Groningen, Groningen, the Netherlands.
authors: Arnold J
authors: AJ
authors: Boersma
authors: Department of Cell Biology, University Medical Center Groningen, University of Groningen, Groningen, the Netherlands.
authors: Inge S
authors: IS
authors: Zuhorn
authors: 1] Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Groningen, the Netherlands. [2] Zernike Institute for Advanced Materials, University of Groningen, Groningen, the Netherlands.
authors: Bert
authors: B
authors: Poolman
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D015374:Biosensing Techniques / Q000379:methods*; D002138:Calibration; D003593:Cytoplasm / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D046911:Macromolecular Substances / Q000032:analysis* / Q000378:metabolism; D057054:Molecular Imaging / Q000379:methods*; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; C485182:Cyan Fluorescent Protein; D008164:Luminescent Proteins; D046911:Macromolecular Substances; D011994:Recombinant Proteins; C054562:yellow fluorescent protein, Bacteria; D049452:Green Fluorescent Proteins
references: Am J Physiol Renal Physiol. 2005 Apr;288(4):F605-13
references: 15753324
references: Biopolymers. 2013 Apr;99(4):239-44
references: 23348671
references: Biochemistry. 2012 Dec 11;51(49):9773-5
references: 23167542
references: J Mol Biol. 1991 Nov 20;222(2):281-300
references: 1960728
references: Biophys J. 2013 Feb 5;104(3):694-704
references: 23442920
references: Annu Rev Biophys. 2008;37:375-97
references: 18573087
references: Cell. 2014 Jan 16;156(1-2):183-94
references: 24361104
references: Phys Rev E Stat Nonlin Soft Matter Phys. 2010 Jun;81(6 Pt 1):061914
references: 20866447
references: Science. 2002 May 3;296(5569):913-6
references: 11988576
references: J Bacteriol. 2006 Sep;188(17):6115-23
references: 16923878
references: Mol Microbiol. 2007 May;64(3):858-71
references: 17462029
references: Annu Rev Biophys. 2008;37:247-63
references: 18573081
references: Antioxid Redox Signal. 2014 Feb 20;20(6):986-99
references: 24205787
references: Cell. 1982 Sep;30(2):345-7
references: 6754085
references: J Am Chem Soc. 2012 Oct 10;134(40):16614-8
references: 22954326
references: Methods Enzymol. 2007;428:487-504
references: 17875435
references: Nat Protoc. 2011 Oct 27;6(11):1806-17
references: 22036883
references: Biophys J. 2004 Nov;87(5):3518-24
references: 15339818
references: Anal Biochem. 1987 Nov 1;166(2):287-97
references: 3434774
references: Chembiochem. 2010 Nov 2;11(16):2264-7
references: 20928879
references: J Bacteriol. 1974 Sep;119(3):736-47
references: 4604283