6ksn: Difference between revisions
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==Structure of a Zn-bound camelid single domain antibody== | |||
<StructureSection load='6ksn' size='340' side='right'caption='[[6ksn]], [[Resolution|resolution]] 2.15Å' scene=''> | |||
== Structural highlights == | |||
<table><tr><td colspan='2'>[[6ksn]] is a 2 chain structure. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=6KSN OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=6KSN FirstGlance]. <br> | |||
</td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=EDO:1,2-ETHANEDIOL'>EDO</scene>, <scene name='pdbligand=NA:SODIUM+ION'>NA</scene>, <scene name='pdbligand=ZN:ZINC+ION'>ZN</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=6ksn FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=6ksn OCA], [http://pdbe.org/6ksn PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=6ksn RCSB], [http://www.ebi.ac.uk/pdbsum/6ksn PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=6ksn ProSAT]</span></td></tr> | |||
</table> | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
Single chain antibodies from camelids have served as powerful tools ranging from diagnostics and therapeutics to crystallization chaperones meant to study protein structure and function. In this study, we isolated a single chain antibody from an Indian dromedary camel (ICab) immunized against a bacterial 14TM helix transporter, NorC, from Staphylococcus aureus We identified this antibody in a yeast display screen built from mononuclear cells isolated from the immunized camel and purified the antibody from E.coli after refolding it from inclusion bodies. The X-ray structure of the antibody at 2.15 A resolution revealed a unique feature within its CDR3 loop, which harbors a Zn(2+)-binding site that substitutes for a loop-stabilizing disulfide bond. We performed mutagenesis to compromise the Zn(2+)-binding site and observed that this change severely hampered antibody stability and its ability to interact with the antigen. The lack of bound Zn(2+)also made the CDR3 loop highly flexible as observed in all-atom simulations. Using confocal imaging of NorC-expressing E.coli spheroplasts, we found that the ICab interacts with the extracellular surface of NorC. This suggests that the ICab could be a valuable tool for detecting methicillin-resistant S.aureus (MRSA) strains that express efflux transporters such as NorC in hospital and community settings. | |||
Isolation and structural characterization of a Zn(2+)-bound single-domain antibody against NorC, a putative multi-drug efflux transporter in bacteria.,Kumar S, Ithayaraja M, Athreya A, Ranjan R, Penmatsa A J Biol Chem. 2019 Nov 7. pii: RA119.010902. doi: 10.1074/jbc.RA119.010902. PMID:31699895<ref>PMID:31699895</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
[[Category: | </div> | ||
<div class="pdbe-citations 6ksn" style="background-color:#fffaf0;"></div> | |||
== References == | |||
<references/> | |||
__TOC__ | |||
</StructureSection> | |||
[[Category: Large Structures]] | |||
[[Category: Athreya, A]] | [[Category: Athreya, A]] | ||
[[Category: Kumar, S]] | [[Category: Kumar, S]] | ||
[[Category: Penmatsa, A]] | [[Category: Penmatsa, A]] | ||
[[Category: Indian camelid single domain antibody]] | |||
[[Category: Metal binding protein]] | |||
[[Category: Vhh]] | |||
