A sensor for quantification of macromolecular crowding in living cells

• Published in: Nature methods Vol. 12; no. 3; pp. 227 - 229
• Main Authors: Boersma, Arnold J, Zuhorn, Inge S, Poolman, Bert
• Format: Journal Article
• Language: English
• Published: New York Nature Publishing Group US 01.03.2015; Nature Publishing Group
• Subjects: 14/19; 14/33; 14/35; 631/1647/1888/2249; 631/1647/338/469; 631/326/41; 631/80/2373; Bacteria; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Bioinformatics; Biological Microscopy; Biological Techniques; Biomedical Engineering/Biotechnology; Biosensing Techniques - methods; Biosensors; brief-communication; Calibration; Cell interaction; Cell research; Cytoplasm; Cytoplasm - metabolism; Energy transfer; Escherichia coli - genetics; Escherichia coli - metabolism; Fluorescence Resonance Energy Transfer; Fluorescence spectroscopy; Green Fluorescent Proteins - genetics; Green Fluorescent Proteins - metabolism; HEK293 Cells; Humans; Life Sciences; Luminescent Proteins - genetics; Luminescent Proteins - metabolism; Macromolecular Substances - analysis; Macromolecular Substances - metabolism; Mammals; Molecular biology; Molecular Imaging - methods; Molecular Sequence Data; Properties; Proteins; Proteomics; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Resonance
• ISSN: 1548-7091; 1548-7105
• DOI: 10.1038/nmeth.3257

A genetically encodable, FRET-based sensor enables quantitative measurements of macromolecular crowding in living bacterial and mammalian cells. 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.