PDBsum entry 3ed8

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protein Protein-protein interface(s) links
Luminescent protein PDB id
Protein chains
229 a.a. *
Waters ×188
* Residue conservation analysis
PDB id:
Name: Luminescent protein
Title: Application of the superfolder yfp bimolecular fluorescence complementation for studying protein-protein interactions i
Structure: Yellow fluorescence protein. Chain: a, b, c, d, e. Engineered: yes. Mutation: yes
Source: Aequorea victoria. Organism_taxid: 6100. Expressed in: escherichia coli. Expression_system_taxid: 562
2.70Å     R-factor:   0.177     R-free:   0.218
Authors: C.Ottmann,M.Weyand
Key ref: C.Ottmann et al. (2009). Applicability of superfolder YFP bimolecular fluorescence complementation in vitro. Biol Chem, 390, 81-90. PubMed id: 19007309
02-Sep-08     Release date:   20-Jan-09    
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Protein chains
Pfam   ArchSchema ?
P42212  (GFP_AEQVI) -  Green fluorescent protein
238 a.a.
229 a.a.*
Key:    PfamA domain  Secondary structure  CATH domain
* PDB and UniProt seqs differ at 23 residue positions (black crosses)

 Gene Ontology (GO) functional annotation 
  GO annot!
  Biological process     generation of precursor metabolites and energy   2 terms 


Biol Chem 390:81-90 (2009)
PubMed id: 19007309  
Applicability of superfolder YFP bimolecular fluorescence complementation in vitro.
C.Ottmann, M.Weyand, A.Wolf, J.Kuhlmann, C.Ottmann.
Bimolecular fluorescence complementation (BiFC) using yellow fluorescent protein (YFP) is a widely employed method to study protein-protein interactions in cells. As yet, this technique has not been used in vitro. To evaluate a possible application of BiFC in vitro, we constructed a 'superfolder split YFP' system where 15 mutations enhance expression of the fusion proteins in Escherichia coli and enable a native purification due to improved solubility. Here, we present the crystal structure of 'superfolder YFP', providing the structural basis for the enhanced folding and stability characteristics. Complementation between the two non-fluorescent YFP fragments fused to HRas and Raf1RBD or to 14-3-3 and PMA2-CT52 resulted in the constitution of the functional fluorophore. The in vivo BiFC with these protein interaction pairs was demonstrated in eukaryotic cell lines as well. Here, we present for the first time BiFC in vitro studies with natively purified superfolder YFP fusion proteins and show the potential and drawbacks of this method for analyzing protein-protein interactions.

Literature references that cite this PDB file's key reference

  PubMed id Reference
21463942 M.Isogai, Y.Kawamoto, K.Inahata, H.Fukada, K.Sugimoto, and T.Tada (2011).
Structure and characteristics of reassembled fluorescent protein, a new insight into the reassembly mechanisms.
  Bioorg Med Chem Lett, 21, 3021-3024.
PDB code: 3ako
20015298 R.H.Rose, S.J.Briddon, and N.D.Holliday (2010).
Bimolecular fluorescence complementation: lighting up seven transmembrane domain receptor signalling networks.
  Br J Pharmacol, 159, 738-750.  
19733184 A.M.Robida, and T.K.Kerppola (2009).
Bimolecular fluorescence complementation analysis of inducible protein interactions: effects of factors affecting protein folding on fluorescent protein fragment association.
  J Mol Biol, 394, 391-409.  
19771338 S.T.Hsu, G.Blaser, and S.E.Jackson (2009).
The folding, stability and conformational dynamics of beta-barrel fluorescent proteins.
  Chem Soc Rev, 38, 2951-2965.  
The most recent references are shown first. Citation data come partly from CiteXplore and partly from an automated harvesting procedure. Note that this is likely to be only a partial list as not all journals are covered by either method. However, we are continually building up the citation data so more and more references will be included with time. Where a reference describes a PDB structure, the PDB code is shown on the right.