7ond
From Proteopedia
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==HaloTag Engineering for Enhanced Fluorogenicity and Kinetics with a Styrylpyridine Dye== | ==HaloTag Engineering for Enhanced Fluorogenicity and Kinetics with a Styrylpyridine Dye== | ||
| - | <StructureSection load='7ond' size='340' side='right'caption='[[7ond]]' scene=''> | + | <StructureSection load='7ond' size='340' side='right'caption='[[7ond]], [[Resolution|resolution]] 1.45Å' scene=''> |
== Structural highlights == | == Structural highlights == | ||
| - | <table><tr><td colspan='2'>Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=7OND OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=7OND FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[7ond]] is a 2 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=7OND OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=7OND FirstGlance]. <br> |
| - | </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=7ond FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=7ond OCA], [https://pdbe.org/7ond PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=7ond RCSB], [https://www.ebi.ac.uk/pdbsum/7ond PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=7ond ProSAT]</span></td></tr> | + | </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=MG:MAGNESIUM+ION'>MG</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='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='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=7ond FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=7ond OCA], [https://pdbe.org/7ond PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=7ond RCSB], [https://www.ebi.ac.uk/pdbsum/7ond PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=7ond 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. | ||
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| + | 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> | ||
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| + | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | ||
| + | </div> | ||
| + | <div class="pdbe-citations 7ond" style="background-color:#fffaf0;"></div> | ||
| + | == References == | ||
| + | <references/> | ||
__TOC__ | __TOC__ | ||
</StructureSection> | </StructureSection> | ||
| + | [[Category: Haloalkane dehalogenase]] | ||
[[Category: Large Structures]] | [[Category: Large Structures]] | ||
| - | [[Category: Liang | + | [[Category: Rhoso]] |
| - | [[Category: Stein A]] | + | [[Category: Liang, A D]] |
| + | [[Category: Stein, A]] | ||
| + | [[Category: Channel dye]] | ||
| + | [[Category: Directed evolution]] | ||
| + | [[Category: Fluorescence]] | ||
| + | [[Category: Fluorescent protein]] | ||
| + | [[Category: Fluorescent reporter]] | ||
| + | [[Category: Halotag]] | ||
| + | [[Category: Imaging]] | ||
| + | [[Category: Mammalian cell]] | ||
Revision as of 20:16, 20 October 2021
HaloTag Engineering for Enhanced Fluorogenicity and Kinetics with a Styrylpyridine Dye
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