5nlu: Difference between revisions
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<StructureSection load='5nlu' size='340' side='right' caption='[[5nlu]], [[Resolution|resolution]] 1.19Å' scene=''> | <StructureSection load='5nlu' size='340' side='right' caption='[[5nlu]], [[Resolution|resolution]] 1.19Å' scene=''> | ||
== Structural highlights == | == Structural highlights == | ||
<table><tr><td colspan='2'>[[5nlu]] is a 1 chain structure. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5NLU OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5NLU FirstGlance]. <br> | <table><tr><td colspan='2'>[[5nlu]] is a 1 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=5NLU OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5NLU FirstGlance]. <br> | ||
</td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=SO4:SULFATE+ION'>SO4</scene></td></tr> | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=SO4:SULFATE+ION'>SO4</scene></td></tr> | ||
<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5nlu FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5nlu OCA], [http://pdbe.org/5nlu PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=5nlu RCSB], [http://www.ebi.ac.uk/pdbsum/5nlu PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=5nlu ProSAT]</span></td></tr> | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5nlu FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5nlu OCA], [http://pdbe.org/5nlu PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=5nlu RCSB], [http://www.ebi.ac.uk/pdbsum/5nlu PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=5nlu ProSAT]</span></td></tr> | ||
</table> | </table> | ||
<div style="background-color:#fffaf0;"> | |||
== 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<ref>PMID:28994409</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
</div> | |||
<div class="pdbe-citations 5nlu" style="background-color:#fffaf0;"></div> | |||
== References == | |||
<references/> | |||
__TOC__ | __TOC__ | ||
</StructureSection> | </StructureSection> | ||
[[Category: Camelus glama]] | |||
[[Category: Andersen, G R]] | [[Category: Andersen, G R]] | ||
[[Category: Andersen, K R]] | [[Category: Andersen, K R]] | ||
Revision as of 09:18, 1 November 2017
Structure of Nb36 crystal form 1Structure of Nb36 crystal form 1
Structural highlights
Publication Abstract from PubMedThe 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
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