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Membrane Biogenesis pp 285–299Cite as

Analysis of Protein Translocation into the Endoplasmic Reticulum of Human Cells

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1033))

Abstract

The development of small-interfering RNA (siRNA)-mediated gene-silencing strategies has made it possible to study the transport of precursors of soluble and membrane proteins into the endoplasmic reticulum (ER) of human cells. In these approaches, a certain target gene is silenced in the cell type of choice, followed by analysis of the effect of this silencing on the biogenesis of a single or set of precursor polypeptide(s) in cell culture or in cell-free assays involving semi-permeabilized cells and in vitro translations systems. These approaches allow for functional analysis of components of the ER-resident protein transport machinery as well as the elucidation of their potential cell-type variations and regulatory mechanisms. The gene-silencing and subsequent plasmid-based complementation carries the additional benefit of facilitating analysis of the consequences of disease-linked mutations in ER transport components.

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Acknowledgment

This work was supported by the Deutsche Forschungsgemeinschaft (DFG).

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Authors and Affiliations

  1. Medical Biochemistry and Molecular Biology, Saarland University, Homburg, Germany

    Johanna Dudek, Sven Lang, Stefan Schorr, Johannes Linxweiler, Markus Greiner & Richard Zimmermann

Authors
  1. Johanna Dudek
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  2. Sven Lang
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  3. Stefan Schorr
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  4. Johannes Linxweiler
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  5. Markus Greiner
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  6. Richard Zimmermann
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Editor information

Editors and Affiliations

  1. Interfaculty Institute of Biochemistry, University of Tuebingen, Tuebingen, Germany

    Doron Rapaport

  2. Cell Biology, University of Kaiserslautern, Kaiserslautern, Germany

    Johannes M. Herrmann

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Dudek, J., Lang, S., Schorr, S., Linxweiler, J., Greiner, M., Zimmermann, R. (2013). Analysis of Protein Translocation into the Endoplasmic Reticulum of Human Cells. In: Rapaport, D., Herrmann, J. (eds) Membrane Biogenesis. Methods in Molecular Biology, vol 1033. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-487-6_18

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  • DOI: https://doi.org/10.1007/978-1-62703-487-6_18

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-486-9

  • Online ISBN: 978-1-62703-487-6

  • eBook Packages: Springer Protocols

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