3K1K

Green fluorescent protein bound to enhancer nanobody

Summary for 3K1K

• Entry DOI: 10.2210/pdb3k1k/pdb
• Related: 3G9A
• Descriptor: Green Fluorescent Protein, Enhancer (3 entities in total)
• Functional Keywords: nanobody, antibody-complex, chromophore, luminescence, photoprotein, luminescent protein-immune system complex, luminescent protein/immune system
• Biological source: Aequorea Victoria; More
• Total number of polymer chains: 4
• Total formula weight: 81553.56

Authors

Kirchhofer, A.,Helma, J.,Schmidthals, K.,Frauer, C.,Cui, S.,Karcher, A.,Pellis, M.,Muyldermans, S.,Delucci, C.C.,Cardoso, M.C.,Leonhardt, H.,Hopfner, K.-P.,Rothbauer, U. (deposition date: 2009-09-28, release date: 2009-12-08, Last modification date: 2024-10-09)

Primary citation

Kirchhofer, A.,Helma, J.,Schmidthals, K.,Frauer, C.,Cui, S.,Karcher, A.,Pellis, M.,Muyldermans, S.,Deulcci, C.C.,Cardoso, M.C.,Leonhardt, H.,Hopfner, K.-P.,Rothbauer, U.
Modulation of protein properties in living cells using nanobodies
Nat.Struct.Mol.Biol., 17:133-138, 2010

PubMed 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.

PubMed: 20010839
DOI: 10.1038/nsmb.1727

Experimental method

X-RAY DIFFRACTION (2.15 Å)