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| | <StructureSection load='5nm0' size='340' side='right'caption='[[5nm0]], [[Resolution|resolution]] 1.50Å' scene=''> | | <StructureSection load='5nm0' size='340' side='right'caption='[[5nm0]], [[Resolution|resolution]] 1.50Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[5nm0]] is a 4 chain structure with sequence from [http://en.wikipedia.org/wiki/Camelus_glama Camelus glama]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5NM0 OCA]. For a <b>guided tour on the structure components</b> use [http://proteopedia.org/fgij/fg.htm?mol=5NM0 FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[5nm0]] is a 4 chain structure with sequence from [https://en.wikipedia.org/wiki/Lama_glama Lama glama]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5NM0 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=5NM0 FirstGlance]. <br> |
| - | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=HG:MERCURY+(II)+ION'>HG</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]] 1.5Å</td></tr> |
| - | <tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[5nlu|5nlu]], [[5nlw|5nlw]]</td></tr> | + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=HG:MERCURY+(II)+ION'>HG</scene></td></tr> |
| - | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://proteopedia.org/fgij/fg.htm?mol=5nm0 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5nm0 OCA], [http://pdbe.org/5nm0 PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=5nm0 RCSB], [http://www.ebi.ac.uk/pdbsum/5nm0 PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=5nm0 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=5nm0 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5nm0 OCA], [https://pdbe.org/5nm0 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=5nm0 RCSB], [https://www.ebi.ac.uk/pdbsum/5nm0 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=5nm0 ProSAT]</span></td></tr> |
| | </table> | | </table> |
| | <div style="background-color:#fffaf0;"> | | <div style="background-color:#fffaf0;"> |
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| | __TOC__ | | __TOC__ |
| | </StructureSection> | | </StructureSection> |
| - | [[Category: Camelus glama]] | + | [[Category: Lama glama]] |
| | [[Category: Large Structures]] | | [[Category: Large Structures]] |
| - | [[Category: Andersen, G R]] | + | [[Category: Andersen GR]] |
| - | [[Category: Andersen, K R]] | + | [[Category: Andersen KR]] |
| - | [[Category: Hansen, S B]] | + | [[Category: Hansen SB]] |
| - | [[Category: Laursen, N S]] | + | [[Category: Laursen NS]] |
| - | [[Category: Ig domain llama single domain antibody nanobody hg derivative]]
| + | |
| - | [[Category: Immune system]]
| + | |
| Structural highlights
Publication Abstract from PubMed
The generation of high-quality protein crystals and the loss of phase information during an X-ray crystallography diffraction experiment represent the major bottlenecks in the determination of novel protein structures. A generic method for introducing Hg atoms into any crystal independent of the presence of free cysteines in the target protein could considerably facilitate the process of obtaining unbiased experimental phases. Nanobodies (single-domain antibodies) have recently been shown to promote the crystallization and structure determination of flexible proteins and complexes. To extend the usability of nanobodies for crystallographic work, variants of the Nb36 nanobody with a single free cysteine at one of four framework-residue positions were developed. These cysteines could be labelled with fluorophores or Hg. For one cysteine variant (Nb36-C85) two nanobody structures were experimentally phased using single-wavelength anomalous dispersion (SAD) and single isomorphous replacement with anomalous signal (SIRAS), taking advantage of radiation-induced changes in Cys-Hg bonding. Importantly, Hg labelling influenced neither the interaction of Nb36 with its antigen complement C5 nor its structure. The results suggest that Cys-Hg-labelled nanobodies may become efficient tools for obtaining de novo phase information during the structure determination of nanobody-protein complexes.
Introducing site-specific cysteines into nanobodies for mercury labelling allows de novo phasing of their crystal structures.,Hansen SB, Laursen NS, Andersen GR, Andersen KR Acta Crystallogr D Struct Biol. 2017 Oct 1;73(Pt 10):804-813. doi:, 10.1107/S2059798317013171. Epub 2017 Sep 27. PMID:28994409[1]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
References
- ↑ Hansen SB, Laursen NS, Andersen GR, Andersen KR. Introducing site-specific cysteines into nanobodies for mercury labelling allows de novo phasing of their crystal structures. Acta Crystallogr D Struct Biol. 2017 Oct 1;73(Pt 10):804-813. doi:, 10.1107/S2059798317013171. Epub 2017 Sep 27. PMID:28994409 doi:http://dx.doi.org/10.1107/S2059798317013171
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