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PDBsum entry 3g9a
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Fluorescent protein/immune system
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PDB id
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3g9a
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References listed in PDB file
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Key reference
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Title
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Modulation of protein properties in living cells using nanobodies.
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Authors
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A.Kirchhofer,
J.Helma,
K.Schmidthals,
C.Frauer,
S.Cui,
A.Karcher,
M.Pellis,
S.Muyldermans,
C.S.Casas-Delucchi,
M.C.Cardoso,
H.Leonhardt,
K.P.Hopfner,
U.Rothbauer.
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Ref.
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Nat Struct Biol, 2010,
17,
133-138.
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PubMed id
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Abstract
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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.
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