6lz2: Difference between revisions
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==Crystal structure of a thermostable green fluorescent protein (TGP) with a synthetic nanobody (Sb44)== | |||
<StructureSection load='6lz2' size='340' side='right'caption='[[6lz2]], [[Resolution|resolution]] 2.03Å' scene=''> | |||
== Structural highlights == | |||
<table><tr><td colspan='2'>[[6lz2]] is a 4 chain structure with sequence from [https://en.wikipedia.org/wiki/Galaxea_fascicularis Galaxea fascicularis] and [https://en.wikipedia.org/wiki/Lama_glama Lama glama]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=6LZ2 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=6LZ2 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]] 2.03Å</td></tr> | |||
<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=ACT:ACETATE+ION'>ACT</scene>, <scene name='pdbligand=CRQ:[2-(3-CARBAMOYL-1-IMINO-PROPYL)-4-(4-HYDROXY-BENZYLIDENE)-5-OXO-4,5-DIHYDRO-IMIDAZOL-1-YL]-ACETIC+ACID'>CRQ</scene>, <scene name='pdbligand=GOL:GLYCEROL'>GOL</scene>, <scene name='pdbligand=NA:SODIUM+ION'>NA</scene>, <scene name='pdbligand=TRS:2-AMINO-2-HYDROXYMETHYL-PROPANE-1,3-DIOL'>TRS</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=6lz2 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=6lz2 OCA], [https://pdbe.org/6lz2 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=6lz2 RCSB], [https://www.ebi.ac.uk/pdbsum/6lz2 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=6lz2 ProSAT]</span></td></tr> | |||
</table> | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
Green fluorescent proteins (GFPs) are widely used to monitor membrane protein expression, purification, and stability. An ideal reporter should be stable itself and provide high sensitivity and yield. Here, we demonstrate that a coral (Galaxea fascicularis) thermostable GFP (TGP) is by such reasons an improved tag compared to the conventional jellyfish GFPs. TGP faithfully reports membrane protein stability at temperatures near 90 degrees C (20-min heating). By contrast, the limit for the two popular GFPs is 64 degrees C and 74 degrees C. Replacing GFPs with TGP increases yield for all four test membrane proteins in four expression systems. To establish TGP as an affinity tag for membrane protein purification, several high-affinity synthetic nanobodies (sybodies), including a non-competing pair, are generated, and the crystal structure of one complex is solved. Given these advantages, we anticipate that TGP becomes a widely used tool for membrane protein structural studies. | |||
An improved fluorescent tag and its nanobodies for membrane protein expression, stability assay, and purification.,Cai H, Yao H, Li T, Hutter CAJ, Li Y, Tang Y, Seeger MA, Li D Commun Biol. 2020 Dec 10;3(1):753. doi: 10.1038/s42003-020-01478-z. PMID:33303987<ref>PMID:33303987</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
[[Category: | </div> | ||
<div class="pdbe-citations 6lz2" style="background-color:#fffaf0;"></div> | |||
==See Also== | |||
*[[Green Fluorescent Protein 3D structures|Green Fluorescent Protein 3D structures]] | |||
== References == | |||
<references/> | |||
__TOC__ | |||
</StructureSection> | |||
[[Category: Galaxea fascicularis]] | |||
[[Category: Lama glama]] | |||
[[Category: Large Structures]] | |||
[[Category: Cai H]] | |||
[[Category: Hutter C]] | |||
[[Category: Li D]] | |||
[[Category: Li T]] | |||
[[Category: Li Y]] | |||
[[Category: Seeger M]] | |||
[[Category: Tang Y]] | |||
[[Category: Yao H]] | |||
Latest revision as of 13:29, 15 November 2023
Crystal structure of a thermostable green fluorescent protein (TGP) with a synthetic nanobody (Sb44)Crystal structure of a thermostable green fluorescent protein (TGP) with a synthetic nanobody (Sb44)
Structural highlights
Publication Abstract from PubMedGreen fluorescent proteins (GFPs) are widely used to monitor membrane protein expression, purification, and stability. An ideal reporter should be stable itself and provide high sensitivity and yield. Here, we demonstrate that a coral (Galaxea fascicularis) thermostable GFP (TGP) is by such reasons an improved tag compared to the conventional jellyfish GFPs. TGP faithfully reports membrane protein stability at temperatures near 90 degrees C (20-min heating). By contrast, the limit for the two popular GFPs is 64 degrees C and 74 degrees C. Replacing GFPs with TGP increases yield for all four test membrane proteins in four expression systems. To establish TGP as an affinity tag for membrane protein purification, several high-affinity synthetic nanobodies (sybodies), including a non-competing pair, are generated, and the crystal structure of one complex is solved. Given these advantages, we anticipate that TGP becomes a widely used tool for membrane protein structural studies. An improved fluorescent tag and its nanobodies for membrane protein expression, stability assay, and purification.,Cai H, Yao H, Li T, Hutter CAJ, Li Y, Tang Y, Seeger MA, Li D Commun Biol. 2020 Dec 10;3(1):753. doi: 10.1038/s42003-020-01478-z. PMID:33303987[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. See AlsoReferences
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