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Fluorescence Anisotropy to Detect In Vivo Stimulus-Induced Changes in Chemoreceptor Packing

  • Vered Frank
  • Ady Vaknin
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1729)

Abstract

The anisotropy of the fluorescence emitted from fluorescent proteins, such as yellow fluorescent protein (YFP), is sensitive to Homo-FRET between the proteins. This effect can be used to detect in vivo ligand-induced changes in packing or conformation of tagged chemoreceptors. Such measurements of clustered or dispersed core-signaling units revealed quantitative dose-dependent responses of these sensors.

Keywords

Fluorescence anisotropy Homo-FRET Chemoreceptors 

Notes

Acknowledgment

The use of fluorescence anisotropy for detecting chemoreceptor responses was pioneered in the lab of Howard C. Berg [5]. This work was funded by Israel Science Foundation.

References

  1. 1.
    Lakowicz JR (2006) Principles of fluorescence spectroscopy, 3rd edn. Springer, New YorkCrossRefGoogle Scholar
  2. 2.
    Clayton AH, Hanley QS, Arndt-Jovin DJ, Subramaniam V, Jovin TM (2002) Dynamic fluorescence anisotropy imaging microscopy in the frequency domain (rFLIM). Biophys J 83:1631–1649CrossRefGoogle Scholar
  3. 3.
    Lidke DS, Nagy P, Barisas BG, Heintzmann R, Post JN et al (2003) Imaging molecular interactions in cells by dynamic and static fluorescence anisotropy (rFLIM and emFRET). Biochem Soc Trans 31(Pt 5):1020–1027CrossRefGoogle Scholar
  4. 4.
    Sharma P, Heo Y, Jang BK, Liu YY, Li JY et al (2004) Nanoscale organization of multiple GPI-anchored proteins in living cell membranes. Cell 116:577–589CrossRefGoogle Scholar
  5. 5.
    Vaknin A, Berg HC (2006) Osmotic stress mechanically perturbs chemoreceptors in Escherichia coli. Proc Natl Acad Sci U S A 103:592–596CrossRefGoogle Scholar
  6. 6.
    Vaknin A, Berg HC (2007) Physical responses of bacterial chemoreceptors. J Mol Biol 366:1416–1423CrossRefGoogle Scholar
  7. 7.
    Frank V, Vaknin A (2013) Prolonged stimuli alter the bacterial chemosensory clusters. Mol Microbiol 88:634–644CrossRefGoogle Scholar
  8. 8.
    Berg HC, Block SM (1984) A miniature flow cell designed for rapid exchange of media under high-power microscope objectives. J Gen Microbiol 130:2915–2920PubMedGoogle Scholar
  9. 9.
    Zacharias DA, Violin JD, Newton AC, Tsien RY et al (2002) Partitioning of lipid-modified monomeric GFPs into membrane microdomains of live cells. Science 296:913–916CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2018

Authors and Affiliations

  1. 1.The Racah Institute of PhysicsThe Hebrew UniversityJerusalemIsrael

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