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Modulation of protein properties in living cells using nanobodies.

Kirchhofer, A. and Helma, J. and Schmidthals, K. and Frauer, C. and Cui, S. and Karcher, A. and Pellis, M. and Muyldermans, S. and Casas-Delucchi, C. S. and Cardoso, M Cristina and Leonhardt, H. and Hopfner, K. P. and Rothbauer, U. (2010):
Modulation of protein properties in living cells using nanobodies.
In: Nature structural & molecular biology, pp. 133-8, 17, (1), ISSN 1545-9985,
[Online-Edition: http://www.cardoso-lab.org/publications/Kirchhofer_2010.pdf],
[Article]

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Abstract

Protein conformation is critically linked to function and often controlled by interactions with regulatory factors. Here we report the selection of camelid-derived single-domain antibodies (nanobodies) that modulate the conformation and spectral properties of the green fluorescent protein (GFP). One nanobody could reversibly reduce GFP fluorescence by a factor of 5, whereas its displacement by a second nanobody caused an increase by a factor of 10. Structural analysis of GFP-nanobody complexes revealed that the two nanobodies induce subtle opposing changes in the chromophore environment, leading to altered absorption properties. Unlike conventional antibodies, the small, stable nanobodies are functional in living cells. Nanobody-induced changes were detected by ratio imaging and used to monitor protein expression and subcellular localization as well as translocation events such as the tamoxifen-induced nuclear localization of estrogen receptor. This work demonstrates that protein conformations can be manipulated and studied with nanobodies in living cells.

Item Type: Article
Erschienen: 2010
Creators: Kirchhofer, A. and Helma, J. and Schmidthals, K. and Frauer, C. and Cui, S. and Karcher, A. and Pellis, M. and Muyldermans, S. and Casas-Delucchi, C. S. and Cardoso, M Cristina and Leonhardt, H. and Hopfner, K. P. and Rothbauer, U.
Title: Modulation of protein properties in living cells using nanobodies.
Language: English
Abstract:

Protein conformation is critically linked to function and often controlled by interactions with regulatory factors. Here we report the selection of camelid-derived single-domain antibodies (nanobodies) that modulate the conformation and spectral properties of the green fluorescent protein (GFP). One nanobody could reversibly reduce GFP fluorescence by a factor of 5, whereas its displacement by a second nanobody caused an increase by a factor of 10. Structural analysis of GFP-nanobody complexes revealed that the two nanobodies induce subtle opposing changes in the chromophore environment, leading to altered absorption properties. Unlike conventional antibodies, the small, stable nanobodies are functional in living cells. Nanobody-induced changes were detected by ratio imaging and used to monitor protein expression and subcellular localization as well as translocation events such as the tamoxifen-induced nuclear localization of estrogen receptor. This work demonstrates that protein conformations can be manipulated and studied with nanobodies in living cells.

Journal or Publication Title: Nature structural & molecular biology
Volume: 17
Number: 1
Divisions: 10 Department of Biology > Cell Biology and Epigenetics
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10 Department of Biology
Date Deposited: 06 Mar 2010 16:27
Official URL: http://www.cardoso-lab.org/publications/Kirchhofer_2010.pdf
Identification Number: doi:10.1038/nsmb.1727
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