4pfe

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Crystal structure of vsfGFP-0Crystal structure of vsfGFP-0

Structural highlights

4pfe is a 2 chain structure. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance.
NonStd Res:
Resources:FirstGlance, OCA, PDBe, RCSB, PDBsum

Function

[GFP_AEQVI] Energy-transfer acceptor. Its role is to transduce the blue chemiluminescence of the protein aequorin into green fluorescent light by energy transfer. Fluoresces in vivo upon receiving energy from the Ca(2+)-activated photoprotein aequorin.

Publication Abstract from PubMed

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 (3x brighter due to dimerization); expression in bacteria and human cells in vivo (1.5x 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.

Rational Structure-Based Design of Bright GFP-Based Complexes with Tunable Dimerization.,Eshaghi M, Sun G, Gruter A, Lim CL, Chee YC, Jung G, Jauch R, Wohland T, Chen SL Angew Chem Int Ed Engl. 2015 Nov 16;54(47):13952-6. doi: 10.1002/anie.201506686. , Epub 2015 Oct 8. PMID:26447926[1]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

References

  1. Eshaghi M, Sun G, Gruter A, Lim CL, Chee YC, Jung G, Jauch R, Wohland T, Chen SL. Rational Structure-Based Design of Bright GFP-Based Complexes with Tunable Dimerization. Angew Chem Int Ed Engl. 2015 Nov 16;54(47):13952-6. doi: 10.1002/anie.201506686. , Epub 2015 Oct 8. PMID:26447926 doi:http://dx.doi.org/10.1002/anie.201506686

4pfe, resolution 2.60Å

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