4PFE

Crystal structure of vsfGFP-0

Summary for 4PFE

• Entry DOI: 10.2210/pdb4pfe/pdb
• Descriptor: Green fluorescent protein (2 entities in total)
• Functional Keywords: beta barrel, fusion protein, homodimer, immune system, fluorescent protein
• Biological source: Aequorea victoria (Jellyfish); More
• Total number of polymer chains: 2
• Total formula weight: 77122.22

Authors

Jauch, R.,Chen, S.L. (deposition date: 2014-04-29, release date: 2015-06-24, Last modification date: 2024-11-06)

Primary citation

Eshaghi, M.,Sun, G.,Gruter, A.,Lim, C.L.,Chee, Y.C.,Jung, G.,Jauch, R.,Wohland, T.,Chen, S.L.
Rational Structure-Based Design of Bright GFP-Based Complexes with Tunable Dimerization.
Angew.Chem.Int.Ed.Engl., 54:13952-13956, 2015

PubMed Abstract: Fluorescent proteins are transformative tools; thus, any brightness increase is a welcome improvement. We invented the "vGFP strategy" based on structural analysis of GFP bound to a single-domain antibody, predicting tunable dimerization, enhanced brightness (ca. 50%), and improved pH resistance. We verified all of these predictions using biochemistry, crystallography, and single-molecule studies. We applied the vsfGFP proteins in three diverse scenarios: single-step immunofluorescence in vitro (3× brighter due to dimerization); expression in bacteria and human cells in vivo (1.5× brighter); and protein fusions showing better pH resistance in human cells in vivo. The vGFP strategy thus allows upgrading of existing applications, is applicable to other fluorescent proteins, and suggests a method for tuning dimerization of arbitrary proteins and optimizing protein properties in general.

PubMed: 26447926
DOI: 10.1002/anie.201506686

Experimental method

X-RAY DIFFRACTION (2.603 Å)