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

Michael R. Stoneman; S. Patowary; M. T. Roesch; D. R. Singh; V. Strogolov; J. A. Oliver; V. Raicu

doi:10.1117/12.875249

11 February 2011 Determination of the stoichiometry, structure, and distribution in living cells of protein complexes from analysis of single-molecular-complexes FRET

Michael R. Stoneman, S. Patowary, M. T. Roesch, D. R. Singh, V. Strogolov, J. A. Oliver, V. Raicu

Author Affiliations +

Proceedings Volume 7903, Multiphoton Microscopy in the Biomedical Sciences XI; 790324 (2011) https://doi.org/10.1117/12.875249
Event: SPIE BiOS, 2011, San Francisco, California, United States

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).

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Michael R. Stoneman, S. Patowary, M. T. Roesch, D. R. Singh, V. Strogolov, J. A. Oliver, and V. Raicu "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 (11 February 2011); https://doi.org/10.1117/12.875249

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