7oo4: Difference between revisions

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<StructureSection load='7oo4' size='340' side='right'caption='[[7oo4]], [[Resolution|resolution]] 2.10&Aring;' scene=''>
<StructureSection load='7oo4' size='340' side='right'caption='[[7oo4]], [[Resolution|resolution]] 2.10&Aring;' scene=''>
== Structural highlights ==
== Structural highlights ==
<table><tr><td colspan='2'>[[7oo4]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Rhoso Rhoso]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=7OO4 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=7OO4 FirstGlance]. <br>
<table><tr><td colspan='2'>Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=7OO4 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=7OO4 FirstGlance]. <br>
</td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=CL:CHLORIDE+ION'>CL</scene>, <scene name='pdbligand=VF2:4-[(E)-2-[1-(7-chloranylheptyl)pyridin-1-ium-4-yl]ethenyl]-N,N-dimethyl-aniline'>VF2</scene></td></tr>
</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 2.1&#8491;</td></tr>
<tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">dhaA ([https://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=1831 RHOSO])</td></tr>
<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=CL:CHLORIDE+ION'>CL</scene>, <scene name='pdbligand=VF2:4-[(E)-2-[1-(7-chloranylheptyl)pyridin-1-ium-4-yl]ethenyl]-N,N-dimethyl-aniline'>VF2</scene></td></tr>
<tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[https://en.wikipedia.org/wiki/Haloalkane_dehalogenase Haloalkane dehalogenase], with EC number [https://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.8.1.5 3.8.1.5] </span></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=7oo4 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=7oo4 OCA], [https://pdbe.org/7oo4 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=7oo4 RCSB], [https://www.ebi.ac.uk/pdbsum/7oo4 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=7oo4 ProSAT]</span></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=7oo4 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=7oo4 OCA], [https://pdbe.org/7oo4 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=7oo4 RCSB], [https://www.ebi.ac.uk/pdbsum/7oo4 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=7oo4 ProSAT]</span></td></tr>
</table>
</table>
<div style="background-color:#fffaf0;">
== Publication Abstract from PubMed ==
HaloTag is a small self-labeling protein that is frequently used for creating fluorescent reporters in living cells. The small-molecule dyes used with HaloTag are almost exclusively based on rhodamine scaffolds, which are often expensive and challenging to synthesize. Herein, we report the engineering of HaloTag for use with a chemically accessible, inexpensive fluorophore based on the dimethylamino-styrylpyridium dye. Through directed evolution, the maximum fluorogenicity and the apparent second-order bioconjugation rate constants could be improved up to 4-fold and 42-fold, respectively. One of the top variants, HT-SP5, enabled reliable imaging in mammalian cells, with a 113-fold fluorescence enhancement over the parent protein. Additionally, crystallographic characterization of selected mutants suggests the chemical origin of the fluorescent enhancement. The improved dye system offers a valuable tool for imaging and illustrates the viability of engineering self-labeling proteins for alternative fluorophores.


HaloTag Engineering for Enhanced Fluorogenicity and Kinetics with a Styrylpyridium Dye.,Miro-Vinyals C, Stein A, Fischer S, Ward TR, Deliz Liang A Chembiochem. 2021 Oct 5. doi: 10.1002/cbic.202100424. PMID:34609782<ref>PMID:34609782</ref>
==See Also==
 
*[[Dehalogenase 3D structures|Dehalogenase 3D structures]]
From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
</div>
<div class="pdbe-citations 7oo4" style="background-color:#fffaf0;"></div>
== References ==
<references/>
__TOC__
__TOC__
</StructureSection>
</StructureSection>
[[Category: Haloalkane dehalogenase]]
[[Category: Large Structures]]
[[Category: Large Structures]]
[[Category: Rhoso]]
[[Category: Liang AD]]
[[Category: Liang, A D]]
[[Category: Stein A]]
[[Category: Stein, A]]
[[Category: Channel dye]]
[[Category: Directed evolution]]
[[Category: Fluorescence]]
[[Category: Fluorescent protein]]
[[Category: Fluorescent reporter]]
[[Category: Halotag]]
[[Category: Imaging]]
[[Category: Mammalian cell]]

Latest revision as of 12:31, 9 October 2024

HaloTag Engineering for Enhanced Fluorogenicity and Kinetics with a Styrylpyridine DyeHaloTag Engineering for Enhanced Fluorogenicity and Kinetics with a Styrylpyridine Dye

Structural highlights

Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance.
Method:X-ray diffraction, Resolution 2.1Å
Ligands:,
Resources:FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT

See Also

7oo4, resolution 2.10Å

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