4kf5: Difference between revisions
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== | ==Crystal Structure of Split GFP complexed with engineered sfCherry with an insertion of GFP fragment== | ||
[[4kf5]] is a 4 chain structure. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4KF5 OCA]. | <StructureSection load='4kf5' size='340' side='right'caption='[[4kf5]], [[Resolution|resolution]] 2.60Å' scene=''> | ||
== Structural highlights == | |||
<table><tr><td colspan='2'>[[4kf5]] is a 4 chain structure with sequence from [https://en.wikipedia.org/wiki/Synthetic_construct Synthetic construct]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4KF5 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=4KF5 FirstGlance]. <br> | |||
</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 2.599Å</td></tr> | |||
<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=CH6:{(4Z)-2-[(1S)-1-AMINO-3-(METHYLSULFANYL)PROPYL]-4-[(4-HYDROXYPHENYL)METHYLIDENE]-5-OXO-4,5-DIHYDRO-1H-IMIDAZOL-1-YL}ACETIC+ACID'>CH6</scene>, <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'>[https://proteopedia.org/fgij/fg.htm?mol=4kf5 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4kf5 OCA], [https://pdbe.org/4kf5 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=4kf5 RCSB], [https://www.ebi.ac.uk/pdbsum/4kf5 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=4kf5 ProSAT]</span></td></tr> | |||
</table> | |||
== Function == | |||
[https://www.uniprot.org/uniprot/GFP_AEQVI 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. | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
A modular strategy for protein crystallization using split green fluorescent protein (GFP) as a crystallization partner is demonstrated. Insertion of a hairpin containing GFP beta-strands 10 and 11 into a surface loop of a target protein provides two chain crossings between the target and the reconstituted GFP compared with the single connection afforded by terminal GFP fusions. This strategy was tested by inserting this hairpin into a loop of another fluorescent protein, sfCherry. The crystal structure of the sfCherry-GFP(10-11) hairpin in complex with GFP(1-9) was determined at a resolution of 2.6 A. Analysis of the complex shows that the reconstituted GFP is attached to the target protein (sfCherry) in a structurally ordered way. This work opens the way to rapidly creating crystallization variants by reconstituting a target protein bearing the GFP(10-11) hairpin with a variety of GFP(1-9) mutants engineered for favorable crystallization. | |||
Split green fluorescent protein as a modular binding partner for protein crystallization.,Nguyen HB, Hung LW, Yeates TO, Terwilliger TC, Waldo GS Acta Crystallogr D Biol Crystallogr. 2013 Dec;69(Pt 12):2513-23. doi:, 10.1107/S0907444913024608. Epub 2013 Nov 19. PMID:24311592<ref>PMID:24311592</ref> | |||
<ref | |||
[[Category: Hung | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | ||
[[Category: Nguyen | </div> | ||
[[Category: Terwilliger | <div class="pdbe-citations 4kf5" style="background-color:#fffaf0;"></div> | ||
[[Category: Waldo | == References == | ||
[[Category: Yeates | <references/> | ||
__TOC__ | |||
</StructureSection> | |||
[[Category: Large Structures]] | |||
[[Category: Synthetic construct]] | |||
[[Category: Hung L-W]] | |||
[[Category: Nguyen HB]] | |||
[[Category: Terwilliger TC]] | |||
[[Category: Waldo GS]] | |||
[[Category: Yeates TO]] | |||
Latest revision as of 14:03, 6 November 2024
Crystal Structure of Split GFP complexed with engineered sfCherry with an insertion of GFP fragmentCrystal Structure of Split GFP complexed with engineered sfCherry with an insertion of GFP fragment
Structural highlights
FunctionGFP_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 PubMedA modular strategy for protein crystallization using split green fluorescent protein (GFP) as a crystallization partner is demonstrated. Insertion of a hairpin containing GFP beta-strands 10 and 11 into a surface loop of a target protein provides two chain crossings between the target and the reconstituted GFP compared with the single connection afforded by terminal GFP fusions. This strategy was tested by inserting this hairpin into a loop of another fluorescent protein, sfCherry. The crystal structure of the sfCherry-GFP(10-11) hairpin in complex with GFP(1-9) was determined at a resolution of 2.6 A. Analysis of the complex shows that the reconstituted GFP is attached to the target protein (sfCherry) in a structurally ordered way. This work opens the way to rapidly creating crystallization variants by reconstituting a target protein bearing the GFP(10-11) hairpin with a variety of GFP(1-9) mutants engineered for favorable crystallization. Split green fluorescent protein as a modular binding partner for protein crystallization.,Nguyen HB, Hung LW, Yeates TO, Terwilliger TC, Waldo GS Acta Crystallogr D Biol Crystallogr. 2013 Dec;69(Pt 12):2513-23. doi:, 10.1107/S0907444913024608. Epub 2013 Nov 19. PMID:24311592[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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