Format

Send to

Choose Destination
Nat Struct Mol Biol. 2010 Jan;17(1):133-8. doi: 10.1038/nsmb.1727. Epub 2009 Dec 13.

Modulation of protein properties in living cells using nanobodies.

Author information

1
Gene Center, Department of Chemistry and Biochemistry, Ludwig-Maximilians University Munich, Munich, Germany.

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.

PMID:
20010839
DOI:
10.1038/nsmb.1727
[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for Nature Publishing Group
Loading ...
Support Center