Author
Listed:
- Jihye Seong
(Neuroscience Program, University of Illinois, Urbana-Champaign, Urbana, Illinois 61801, USA.)
- Mingxing Ouyang
(University of Illinois, Urbana-Champaign, Urbana, Illinois 61801, USA.)
- Taejin Kim
(Neuroscience Program, University of Illinois, Urbana-Champaign, Urbana, Illinois 61801, USA.)
- Jie Sun
(University of Illinois, Urbana-Champaign, Urbana, Illinois 61801, USA.)
- Po-Chao Wen
(Center for Biophysics and Computational Biology, University of Illinois, Urbana-Champaign, Urbana, Illinois 61801, USA.)
- Shaoying Lu
(University of Illinois, Urbana-Champaign, Urbana, Illinois 61801, USA.)
- Yue Zhuo
(University of Illinois, Urbana-Champaign, Urbana, Illinois 61801, USA.)
- Nicholas M. Llewellyn
(University of Illinois, Urbana-Champaign, Urbana, Illinois 61801, USA.)
- David D. Schlaepfer
(Moores Cancer Center, University of California, San Diego, La Jolla, California 92093, USA.)
- Jun-Lin Guan
(University of Michigan Medical School, Ann Arbor, Michigan 48109, USA.)
- Shu Chien
(University of California, San Diego, La Jolla, California 92093, USA.)
- Yingxiao Wang
(Neuroscience Program, University of Illinois, Urbana-Champaign, Urbana, Illinois 61801, USA.
University of Illinois, Urbana-Champaign, Urbana, Illinois 61801, USA.
University of Illinois, Urbana-Champaign, Urbana, Illinois 61801, USA.
Center for Biophysics and Computational Biology, University of Illinois, Urbana-Champaign, Urbana, Illinois 61801, USA.)
Abstract
Proper subcellular localization of focal adhesion kinase (FAK) is crucial for many cellular processes. It remains, however, unclear how FAK activity is regulated at subcellular compartments. To visualize the FAK activity at different membrane microdomains, we develop a fluorescence resonance energy transfer (FRET)-based FAK biosensor, and target it into or outside of detergent-resistant membrane (DRM) regions at the plasma membrane. Here we show that, on cell adhesion to extracellular matrix proteins or stimulation by platelet-derived growth factor (PDGF), the FRET responses of DRM-targeting FAK biosensor are stronger than that at non-DRM regions, suggesting that FAK activation can occur at DRM microdomains. Further experiments reveal that the PDGF-induced FAK activation is mediated and maintained by Src activity, whereas FAK activation on cell adhesion is independent of, and in fact essential for the Src activation. Therefore, FAK is activated at membrane microdomains with distinct activation mechanisms in response to different physiological stimuli.
Suggested Citation
Jihye Seong & Mingxing Ouyang & Taejin Kim & Jie Sun & Po-Chao Wen & Shaoying Lu & Yue Zhuo & Nicholas M. Llewellyn & David D. Schlaepfer & Jun-Lin Guan & Shu Chien & Yingxiao Wang, 2011.
"Detection of focal adhesion kinase activation at membrane microdomains by fluorescence resonance energy transfer,"
Nature Communications, Nature, vol. 2(1), pages 1-9, September.
Handle:
RePEc:nat:natcom:v:2:y:2011:i:1:d:10.1038_ncomms1414
DOI: 10.1038/ncomms1414
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