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Exploring Protein Superstructures and Dynamics in Live Bacterial Cells Using Single-Molecule and Superresolution Imaging

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Single Molecule Analysis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 783))

Abstract

Single-molecule imaging enables biophysical measurements devoid of ensemble averaging, gives enhanced spatial resolution beyond the optical diffraction limit, and enables superresolution reconstruction of structures beyond the diffraction limit. This work summarizes how single-molecule and superresolution imaging can be applied to the study of protein dynamics and superstructures in live Caulobacter crescentus cells to illustrate the power of these methods in bacterial imaging. Based on these techniques, the diffusion coefficient and dynamics of the histidine protein kinase PleC, the localization behavior of the polar protein PopZ, and the treadmilling behavior and protein superstructure of the structural protein MreB are investigated with sub-40-nm spatial resolution, all in live cells.

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Acknowledgments

The authors warmly thank members of the Shapiro laboratory, specifically Ellen Judd, Harley McAdams, Grant Bowman, and Zemer Gitai, as well as members of the Moerner laboratory, specifically, Jason Deich, Marcelle Koenig, So Yeon Kim, Anika Kinkhabwala, Michael Thompson, and Nicole Tselentis, for their contributions to this work. This research was supported in part by DARPA Grant MDA-972-00-1-0032, NSF Grant MCB-0212503, Department of Energy Grants DE-FG02-04ER63777 and DE_FG02-05ER64136 9 (to LS), and NIH Grants P20-HG003638, R01-GM086196, R01-GM085437, R01-GM51426 (to LS), and R01-GM32506 (to LS).

Author information

Authors and Affiliations

  1. Department of Chemistry, University of Michigan, Ann Arbor, MI, USA

    Julie S. Biteen

  2. Department of Developmental Biology, Stanford University, Stanford, CA, USA

    Lucy Shapiro

  3. Department of Chemistry, Stanford University, Stanford, CA, USA

    W. E. Moerner

Authors
  1. Julie S. Biteen
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  2. Lucy Shapiro
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  3. W. E. Moerner
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Corresponding author

Correspondence to Julie S. Biteen .

Editor information

Editors and Affiliations

  1. Dept. Physics & Astronomy, Section Physics of Complex Systems, Vrije Universiteit Amsterdam, De Boelelaan 1081, Amsterdam, 1081 HV, Netherlands

    Erwin J. G. Peterman

  2. Dept. Physics & Astronomy, Section Physics of Complex Systems, Vrije Universiteit Amsterdam, De Boelelaan 1081, Amsterdam, 1081 HV, Netherlands

    Gijs J. L. Wuite

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Biteen, J.S., Shapiro, L., Moerner, W.E. (2011). Exploring Protein Superstructures and Dynamics in Live Bacterial Cells Using Single-Molecule and Superresolution Imaging. In: Peterman, E., Wuite, G. (eds) Single Molecule Analysis. Methods in Molecular Biology, vol 783. Humana Press. https://doi.org/10.1007/978-1-61779-282-3_8

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  • DOI: https://doi.org/10.1007/978-1-61779-282-3_8

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-281-6

  • Online ISBN: 978-1-61779-282-3

  • eBook Packages: Springer Protocols

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