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Proceedings Article

Determination of the stoichiometry, structure, and distribution in living cells of protein complexes from analysis of single-molecular-complexes FRET

[+] Author Affiliations
Michael R. Stoneman, S. Patowary, M. T. Roesch, D. R. Singh, V. Strogolov, J. A. Oliver, V. Raicu

Univ. of Wisconsin-Milwaukee (USA)

Proc. SPIE 7903, Multiphoton Microscopy in the Biomedical Sciences XI, 790324 (February 10, 2011); doi:10.1117/12.875249
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From Conference Volume 7903

  • Multiphoton Microscopy in the Biomedical Sciences XI
  • Ammasi Periasamy; Karsten König; Peter T. C. So
  • San Francisco, California, USA | January 22, 2011

abstract

Advances in two-photon microscopy with spectral resolution (TPM-SR) and the development of a simple theory of Förster Resonance Energy Transfer (FRET) for single molecular complexes recently lead to the development of a novel method for the determination of structure and localization in living cells of membrane protein complexes (Raicu et al., Nature Photon. , 3, 2009). An appealing feature of this method is its ability to provide such important information while being unaffected by spurious signals originating from stochastic FRET (Singh and Raicu, Biophys. J. , 98, 2010). We will present the results obtained from our recent studies of trimeric FRET calibration standards expressed in the cytoplasm of Chinese hamster ovary (CHO) cells, as well as a model G protein-coupled receptor expressed in the membrane of yeast. Emphasis will be placed on the measurement and analysis of single-molecular-complex FRET data for determination of the quaternary structure of some proteins (or the protein complex structure).

© (2011) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
Citation

Michael R. Stoneman ; S. Patowary ; M. T. Roesch ; D. R. Singh ; V. Strogolov, et al.
"Determination of the stoichiometry, structure, and distribution in living cells of protein complexes from analysis of single-molecular-complexes FRET", Proc. SPIE 7903, Multiphoton Microscopy in the Biomedical Sciences XI, 790324 (February 10, 2011); doi:10.1117/12.875249; http://dx.doi.org/10.1117/12.875249


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