6al0: Difference between revisions
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The | ==The NZ-1 Fab complexed with the PDZ tandem fragment of A. aeolicus S2P homolog with the PA12 tag inserted between the residues 263 and 267== | ||
<StructureSection load='6al0' size='340' side='right' caption='[[6al0]], [[Resolution|resolution]] 2.60Å' scene=''> | |||
== Structural highlights == | |||
<table><tr><td colspan='2'>[[6al0]] is a 3 chain structure. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=6AL0 OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=6AL0 FirstGlance]. <br> | |||
</td></tr><tr id='NonStdRes'><td class="sblockLbl"><b>[[Non-Standard_Residue|NonStd Res:]]</b></td><td class="sblockDat"><scene name='pdbligand=PCA:PYROGLUTAMIC+ACID'>PCA</scene>, <scene name='pdbligand=SNN:L-3-AMINOSUCCINIMIDE'>SNN</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=6al0 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=6al0 OCA], [http://pdbe.org/6al0 PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=6al0 RCSB], [http://www.ebi.ac.uk/pdbsum/6al0 PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=6al0 ProSAT]</span></td></tr> | |||
</table> | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
An antibody fragment that recognizes the tertiary structure of a target protein with high affinity can be utilized as a crystallization chaperone. Difficulties in establishing conformation-specific antibodies, however, limit the applicability of antibody fragment-assisted crystallization. Here, we attempted to establish an alternative method to promote the crystallization of target proteins using an already established anti-tag antibody. The monoclonal antibody NZ-1 recognizes the PA tag with an extremely high affinity. It was also established that the PA tag is accommodated in the antigen-binding pocket in a bent conformation, compatible with an insertion into loop regions on the target. We, therefore, explored the application of NZ-1 Fab as a crystallization chaperone that complexes with a target protein displaying a PA tag. Specifically, we inserted the PA tag into the beta-hairpins of the PDZ tandem fragment of a bacterial Site-2 protease. We crystallized the PA-inserted PDZ tandem mutants with the NZ-1 Fab and solved the co-crystal structure to analyze their interaction modes. Although the initial insertion designs produced only moderate-resolution structures, eliminating the solvent-accessible space between the NZ-1 Fab and target PDZ tandem improved the diffraction qualities remarkably. Our results demonstrate that the NZ-1-PA system efficiently promotes crystallization of the target protein. The present work also suggests that beta-hairpins are suitable sites for the PA insertion because the PA tag contains a Pro-Gly sequence with a propensity for a beta-turn conformation. | |||
Application of the NZ-1 Fab as a crystallization chaperone for PA tag-inserted target proteins.,Tamura R, Oi R, Akashi S, Kaneko MK, Kato Y, Nogi T Protein Sci. 2019 Jan 21. doi: 10.1002/pro.3580. PMID:30666745<ref>PMID:30666745</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
[[Category: | </div> | ||
<div class="pdbe-citations 6al0" style="background-color:#fffaf0;"></div> | |||
== References == | |||
<references/> | |||
__TOC__ | |||
</StructureSection> | |||
[[Category: Kaneko, M K]] | |||
[[Category: Kato, Y]] | |||
[[Category: Nogi, T]] | |||
[[Category: Oi, R]] | [[Category: Oi, R]] | ||
[[Category: Tamura, R]] | [[Category: Tamura, R]] | ||
[[Category: Protease]] | |||
[[Category: Signaling protein]] | |||