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3ed8
From Proteopedia
Revision as of 12:13, 29 September 2014 by OCA (Talk | contribs)
3ed8 is a 5 chain structure with sequence from Aequorea victoria. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance.
Check, as determined by ConSurfDB. You may read the explanation of the method and the full data available from ConSurf.
Publication Abstract from PubMed
Abstract 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.
Applicability of superfolder YFP bimolecular fluorescence complementation in vitro.,Ottmann C, Weyand M, Wolf A, Kuhlmann J, Ottmann C Biol Chem. 2009 Jan;390(1):81-90. PMID:19007309[1]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
↑ Ottmann C, Weyand M, Wolf A, Kuhlmann J, Ottmann C. Applicability of superfolder YFP bimolecular fluorescence complementation in vitro. Biol Chem. 2009 Jan;390(1):81-90. PMID:19007309 doi:10.1515/BC.2009.008
