3OGO

Structure of the GFP:GFP-nanobody complex at 2.8 A resolution in spacegroup P21212

Replaces:  3MIQ

Summary for 3OGO

• Entry DOI: 10.2210/pdb3ogo/pdb
• Descriptor: Green fluorescent protein, GFP-nanobody, ISOPROPYL ALCOHOL, ... (4 entities in total)
• Functional Keywords: gfp, gfp-nanobody, beta-barrel, antibody, luminescent protein-immune system complex, fluorescent protein-immune system complex, fluorescent protein/immune system
• Biological source: Aequorea victoria (Jellyfish); More
• Total number of polymer chains: 8
• Total formula weight: 167800.02

Authors

Kubala, M.H.,Kovtun, O.,Alexandrov, K.,Collins, B.M. (deposition date: 2010-08-17, release date: 2010-08-25, Last modification date: 2024-11-06)

Primary citation

Kubala, M.H.,Kovtun, O.,Alexandrov, K.,Collins, B.M.
Structural and thermodynamic analysis of the GFP:GFP-nanobody complex.
Protein Sci., 19:2389-2401, 2010

PubMed Abstract: The green fluorescent protein (GFP)-nanobody is a single-chain VHH antibody domain developed with specific binding activity against GFP and is emerging as a powerful tool for isolation and cellular engineering of fluorescent protein fusions in many different fields of biological research. Using X-ray crystallography and isothermal titration calorimetry, we determine the molecular details of GFP:GFP-nanobody complex formation and explain the basis of high affinity and at the same time high specificity of protein binding. Although the GFP-nanobody can also bind YFP, it cannot bind the closely related CFP or other fluorescent proteins from the mFruit series. CFP differs from GFP only within the central chromophore and at one surface amino acid position, which lies in the binding interface. Using this information, we have engineered a CFP variant (I146N) that is also able to bind the GFP-nanobody with high affinity, thus extending the toolbox of genetically encoded fluorescent probes that can be isolated using the GFP-nanobody.

PubMed: 20945358
DOI: 10.1002/pro.519

Experimental method

X-RAY DIFFRACTION (2.8 Å)