Modulation of protein properties in living cells using nanobodies.

TitleModulation of protein properties in living cells using nanobodies.
Publication TypeJournal Article
Year of Publication2010
AuthorsKirchhofer, A., Helma J., Schmidthals K., Frauer C., Cui S., Karcher A., Pellis M., Muyldermans S., Casas-Delucchi C. S., M Cardoso C., Leonhardt H., Hopfner K-P., and Rothbauer U.
JournalNat Struct Mol Biol
Volume17
Issue1
Pagination133-8
Date Published2010 Jan
ISSN1545-9985
KeywordsAnimals, Antibodies, Camels, Crystallization, Fluorescence, Green Fluorescent Proteins, Models, Molecular, Protein Conformation, Spectrometry, Fluorescence
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.

DOI10.1038/nsmb.1727
Alternate JournalNat. Struct. Mol. Biol.
PubMed ID20010839
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