3ed8: Difference between revisions
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==Application of the superfolder YFP bimolecular fluorescence complementation for studying protein-protein interactions in vitro== | ==Application of the superfolder YFP bimolecular fluorescence complementation for studying protein-protein interactions in vitro== | ||
<StructureSection load='3ed8' size='340' side='right' caption='[[3ed8]], [[Resolution|resolution]] 2.70Å' scene=''> | <StructureSection load='3ed8' size='340' side='right'caption='[[3ed8]], [[Resolution|resolution]] 2.70Å' scene=''> | ||
== Structural highlights == | == Structural highlights == | ||
<table><tr><td colspan='2'>[[3ed8]] is a 5 chain structure with sequence from [http://en.wikipedia.org/wiki/Aeqvi Aeqvi]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=3ED8 OCA]. For a <b>guided tour on the structure components</b> use [http:// | <table><tr><td colspan='2'>[[3ed8]] is a 5 chain structure with sequence from [http://en.wikipedia.org/wiki/Aeqvi Aeqvi]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=3ED8 OCA]. For a <b>guided tour on the structure components</b> use [http://proteopedia.org/fgij/fg.htm?mol=3ED8 FirstGlance]. <br> | ||
</td></tr><tr id='NonStdRes'><td class="sblockLbl"><b>[[Non-Standard_Residue|NonStd Res:]]</b></td><td class="sblockDat"><scene name='pdbligand=CRO:{2-[(1R,2R)-1-AMINO-2-HYDROXYPROPYL]-4-(4-HYDROXYBENZYLIDENE)-5-OXO-4,5-DIHYDRO-1H-IMIDAZOL-1-YL}ACETIC+ACID'>CRO</scene></td></tr> | </td></tr><tr id='NonStdRes'><td class="sblockLbl"><b>[[Non-Standard_Residue|NonStd Res:]]</b></td><td class="sblockDat"><scene name='pdbligand=CRO:{2-[(1R,2R)-1-AMINO-2-HYDROXYPROPYL]-4-(4-HYDROXYBENZYLIDENE)-5-OXO-4,5-DIHYDRO-1H-IMIDAZOL-1-YL}ACETIC+ACID'>CRO</scene></td></tr> | ||
<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http:// | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://proteopedia.org/fgij/fg.htm?mol=3ed8 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=3ed8 OCA], [http://pdbe.org/3ed8 PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=3ed8 RCSB], [http://www.ebi.ac.uk/pdbsum/3ed8 PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=3ed8 ProSAT]</span></td></tr> | ||
</table> | </table> | ||
== Evolutionary Conservation == | == Evolutionary Conservation == | ||
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==See Also== | ==See Also== | ||
*[[Green Fluorescent Protein|Green Fluorescent Protein]] | *[[Green Fluorescent Protein 3D structures|Green Fluorescent Protein 3D structures]] | ||
== References == | == References == | ||
<references/> | <references/> | ||
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</StructureSection> | </StructureSection> | ||
[[Category: Aeqvi]] | [[Category: Aeqvi]] | ||
[[Category: Large Structures]] | |||
[[Category: Ottmann, C]] | [[Category: Ottmann, C]] | ||
[[Category: Weyand, M]] | [[Category: Weyand, M]] | ||
Revision as of 13:22, 9 September 2020
Application of the superfolder YFP bimolecular fluorescence complementation for studying protein-protein interactions in vitroApplication of the superfolder YFP bimolecular fluorescence complementation for studying protein-protein interactions in vitro
Structural highlights
Evolutionary Conservation Check, as determined by ConSurfDB. You may read the explanation of the method and the full data available from ConSurf. Publication Abstract from PubMedAbstract 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. See AlsoReferences
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