Histochemistry and Cell Biology

, Volume 142, Issue 1, pp 61–67 | Cite as

Spatial distribution and structural arrangement of a murine cytomegalovirus glycoprotein detected by SPDM localization microscopy

  • Patrick Müller
  • Niels A. Lemmermann
  • Rainer Kaufmann
  • Manuel Gunkel
  • Daniel Paech
  • Georg Hildenbrand
  • Rafaela Holtappels
  • Christoph Cremer
  • Michael Hausmann
Original Paper

Abstract

Novel approaches of localization microscopy have opened new insights into the molecular nano-cosmos of cells. We applied a special embodiment called spectral position determination microscopy (SPDM) that has the advantage to run with standard fluorescent dyes or proteins under standard preparation conditions. Pointillist images with a resolution in the order of 10 nm can be obtained by SPDM. Therefore, vector pEYFP-m164, encoding the murine cytomegalovirus glycoprotein gp36.5/m164 fused to enhanced yellow fluorescent protein, was transiently transfected into COS-7 cells. This protein shows exceptional intracellular trafficking dynamics, moving within the endoplasmic reticulum (ER) and outer nuclear membrane. The molecular positions of gp36.5/m164 were visualized and determined by SPDM imaging. From the position point patterns of the protein molecules, their arrangements were quantified by next neighbour distance analyses. Three different structural arrangements were discriminated: (a) a linear distribution along the membrane, (b) a highly structured distribution in the ER, and (c) a homogenous distribution in the cellular cytoplasm. The results indicate that the analysis of next neighbour distances on the nano-scale allows the identification and discrimination of different structural arrangements of molecules within their natural cellular environment.

Keywords

Spectral position determination microscopy Single molecule localization microscopy Super-resolution gp36.5/m164 glycoprotein Murine cytomegalovirus Nano-distance measurements Spatial structures 

Notes

Acknowledgments

The authors gratefully acknowledge the financial support of the Deutsche Forschungsgemeinschaft (German Research Council) to C.C., and of the Bundesministerium für Bildung und Forschung (Federal Ministry for Education and Research) to M.H., R.H. was supported by the Deutsche Forschungsgemeinschaft, Collaborative Research Centre (SFB) 490 (TP-E3) and the intramural funding in program IFF-I of the University Medical Center, Mainz, Germany. N.A.L. received intramural funding in the young investigator program MAIFOR of the University Medical Center, Mainz, Germany. The authors thank Udo Birk, IMB, Mainz, for technical contributions in the development of the SPDM instrumentation.

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Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Patrick Müller
    • 1
  • Niels A. Lemmermann
    • 2
  • Rainer Kaufmann
    • 1
    • 6
  • Manuel Gunkel
    • 1
    • 7
  • Daniel Paech
    • 1
    • 8
  • Georg Hildenbrand
    • 1
    • 3
  • Rafaela Holtappels
    • 2
  • Christoph Cremer
    • 1
    • 4
    • 5
  • Michael Hausmann
    • 1
  1. 1.Kirchhoff-Institute for PhysicsHeidelbergGermany
  2. 2.Institute for VirologyUniversity Medical Center MainzMainzGermany
  3. 3.Department of Radiation OncologyUniversity Medical Center MannheimMannheimGermany
  4. 4.Institute for Pharmacy and Molecular BiotechnologyUniversity HeidelbergHeidelbergGermany
  5. 5.Institute of Molecular Biology (IMB)MainzGermany
  6. 6.Division of Structural Biology, Wellcome Trust Centre for Human Genetics and Department of BiochemistryUniversity of OxfordOxfordUK
  7. 7.BioquantUniversity of HeidelbergHeidelbergGermany
  8. 8.Division of NeuroradiologyUniversity Medical Center HeidelbergHeidelbergGermany

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