6y7b: Difference between revisions

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<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=6y7b FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=6y7b OCA], [https://pdbe.org/6y7b PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=6y7b RCSB], [https://www.ebi.ac.uk/pdbsum/6y7b PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=6y7b 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=6y7b FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=6y7b OCA], [https://pdbe.org/6y7b PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=6y7b RCSB], [https://www.ebi.ac.uk/pdbsum/6y7b PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=6y7b ProSAT]</span></td></tr>
</table>
</table>
<div style="background-color:#fffaf0;">
== Publication Abstract from PubMed ==
The self-labeling protein tags (SLPs) HaloTag7, SNAP-tag, and CLIP-tag allow the covalent labeling of fusion proteins with synthetic molecules for applications in bioimaging and biotechnology. To guide the selection of an SLP-substrate pair and provide guidelines for the design of substrates, we report a systematic and comparative study of the labeling kinetics and substrate specificities of HaloTag7, SNAP-tag, and CLIP-tag. HaloTag7 reaches almost diffusion-limited labeling rate constants with certain rhodamine substrates, which are more than 2 orders of magnitude higher than those of SNAP-tag for the corresponding substrates. SNAP-tag labeling rate constants, however, are less affected by the structure of the label than those of HaloTag7, which vary over 6 orders of magnitude for commonly employed substrates. Determining the crystal structures of HaloTag7 and SNAP-tag labeled with fluorescent substrates allowed us to rationalize their substrate preferences. We also demonstrate how these insights can be exploited to design substrates with improved labeling kinetics.
Kinetic and Structural Characterization of the Self-Labeling Protein Tags HaloTag7, SNAP-tag, and CLIP-tag.,Wilhelm J, Kuhn S, Tarnawski M, Gotthard G, Tunnermann J, Tanzer T, Karpenko J, Mertes N, Xue L, Uhrig U, Reinstein J, Hiblot J, Johnsson K Biochemistry. 2021 Aug 24;60(33):2560-2575. doi: 10.1021/acs.biochem.1c00258., Epub 2021 Aug 2. PMID:34339177<ref>PMID:34339177</ref>
From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
</div>
<div class="pdbe-citations 6y7b" style="background-color:#fffaf0;"></div>


==See Also==
==See Also==
*[[Dehalogenase 3D structures|Dehalogenase 3D structures]]
*[[Dehalogenase 3D structures|Dehalogenase 3D structures]]
== References ==
<references/>
__TOC__
__TOC__
</StructureSection>
</StructureSection>

Revision as of 10:56, 10 November 2021

X-ray structure of the Haloalkane dehalogenase HaloTag7 labeled with a chloroalkane-carbopyronine fluorophore substrateX-ray structure of the Haloalkane dehalogenase HaloTag7 labeled with a chloroalkane-carbopyronine fluorophore substrate

Structural highlights

6y7b is a 5 chain structure with sequence from Rhoso. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance.
Ligands:,
Gene:dhaA (RHOSO)
Activity:Haloalkane dehalogenase, with EC number 3.8.1.5
Resources:FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT

Publication Abstract from PubMed

The self-labeling protein tags (SLPs) HaloTag7, SNAP-tag, and CLIP-tag allow the covalent labeling of fusion proteins with synthetic molecules for applications in bioimaging and biotechnology. To guide the selection of an SLP-substrate pair and provide guidelines for the design of substrates, we report a systematic and comparative study of the labeling kinetics and substrate specificities of HaloTag7, SNAP-tag, and CLIP-tag. HaloTag7 reaches almost diffusion-limited labeling rate constants with certain rhodamine substrates, which are more than 2 orders of magnitude higher than those of SNAP-tag for the corresponding substrates. SNAP-tag labeling rate constants, however, are less affected by the structure of the label than those of HaloTag7, which vary over 6 orders of magnitude for commonly employed substrates. Determining the crystal structures of HaloTag7 and SNAP-tag labeled with fluorescent substrates allowed us to rationalize their substrate preferences. We also demonstrate how these insights can be exploited to design substrates with improved labeling kinetics.

Kinetic and Structural Characterization of the Self-Labeling Protein Tags HaloTag7, SNAP-tag, and CLIP-tag.,Wilhelm J, Kuhn S, Tarnawski M, Gotthard G, Tunnermann J, Tanzer T, Karpenko J, Mertes N, Xue L, Uhrig U, Reinstein J, Hiblot J, Johnsson K Biochemistry. 2021 Aug 24;60(33):2560-2575. doi: 10.1021/acs.biochem.1c00258., Epub 2021 Aug 2. PMID:34339177[1]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

See Also

References

  1. Wilhelm J, Kuhn S, Tarnawski M, Gotthard G, Tunnermann J, Tanzer T, Karpenko J, Mertes N, Xue L, Uhrig U, Reinstein J, Hiblot J, Johnsson K. Kinetic and Structural Characterization of the Self-Labeling Protein Tags HaloTag7, SNAP-tag, and CLIP-tag. Biochemistry. 2021 Aug 24;60(33):2560-2575. doi: 10.1021/acs.biochem.1c00258., Epub 2021 Aug 2. PMID:34339177 doi:http://dx.doi.org/10.1021/acs.biochem.1c00258

6y7b, resolution 3.10Å

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