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Spatio-Temporal Manipulation of Small GTPase Activity at Subcellular Level and on Timescale of Seconds in Living Cells
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Bioengineering
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JoVE Journal Bioengineering
Spatio-Temporal Manipulation of Small GTPase Activity at Subcellular Level and on Timescale of Seconds in Living Cells

Spatio-Temporal Manipulation of Small GTPase Activity at Subcellular Level and on Timescale of Seconds in Living Cells

DOI:
10.3791/3794-v

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10:27 min

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March 09, 2012

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, , , , , ,
1Department of Cell Biology, Center for Cell Dynamics,Johns Hopkins University, 2Graduate School of Pharmaceutical Sciences,University of Tokyo, 3Biomedical Engineering,Johns Hopkins University

Chapters

  • 00:05Title
  • 02:17Transfection of Plasmid DNA
  • 04:02Preparation of cRb-A Solution
  • 05:22Microscopy and Light Illumination at the Subcellular Level
  • 07:07Examining the Effect of RAC Activation on Local Ruffle Formation
  • 09:29Conclusion

Summary

Automatic Translation

Automatic Translation

A method for spatio-temporal control of small GTPase activity by light is described. This method is based on rapamycin-induced FKBP-FRB heterodimerization and photo-caging systems. Optimization of light-irradiation enables the spatio-temporally controlled activation of small GTPases at the subcellular level.

Tags

Spatio-temporal ManipulationSmall GTPase ActivitySubcellular LevelSecondsLiving CellsDynamic RegulationRho FamilyGuanosine TriphosphatasesCellular FunctionsVisualizationReal TimePerturbationProtein ActivitiesSubcellular LocationTimingLight-induced ActivationRapamycin-induced FKBP-FRB HeterodimerizationPhoto-caging MethodRapamycinRapid Inducible ActivationInactivationTranslocationPlasma MembraneFRB AnchorPhoto-caging SystemRapamycin Analogs

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