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4bjh

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Revision as of 11:43, 4 August 2016

Crystal Structure of the Aquifex Reactor Complex Formed by Dihydroorotase (H180A, H232A) with Dihydroorotate and Aspartate Transcarbamoylase with N-(phosphonacetyl)-L-aspartate (PALA)

Structural highlights

4bjh is a 2 chain structure with sequence from "aquifex_aeolicus"_huber_and_stetter_2001 "aquifex aeolicus" huber and stetter 2001. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance.
Ligands:	, , , , ,
Activity:	Dihydroorotase, with EC number 3.5.2.3
Resources:	FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT

Publication Abstract from PubMed

BACKGROUND: Dihydroorotase (DHO) is a zinc metalloenzyme, although the number of active site zinc ions has been controversial. E. coli DHO was initially thought to have a mononuclear metal center, but the subsequent X-ray structure clearly showed two zinc ions, alpha and beta, at the catalytic site. Aquifex aeolicus DHO, is a dodecamer comprised of six DHO and six aspartate transcarbamoylase (ATC) subunits. The isolated DHO monomer, which lacks catalytic activity, has an intact alpha-site and conserved beta-site ligands, but the geometry of the second metal binding site is completely disrupted. However, the putative beta-site is restored when the complex with ATC is formed and DHO activity is regained. Nevertheless, the X-ray structure of the complex revealed a single zinc ion at the active site. The structure of DHO from the pathogenic organism, S. aureus showed that it also has a single active site metal ion. RESULTS: Zinc analysis showed that the enzyme has one zinc/DHO subunit and the addition of excess metal ion did not stimulate catalytic activity, nor alter the kinetic parameters. The metal free apoenzyme was inactive, but the full activity was restored upon the addition of one equivalent of Zn2+ or Co2+. Moreover, deletion of the beta-site by replacing the His180 and His232 with alanine had no effect on catalysis in the presence or absence of excess zinc. The 2.2 A structure of the double mutant confirmed that the beta-site was eliminated but that the active site remained otherwise intact. CONCLUSIONS: Thus, kinetically competent A. aeolicus DHO has a mononuclear metal center. In contrast, elimination of the putative second metal binding site in amidohydrolyases with a binuclear metal center, resulted in the abolition of catalytic activity. The number of active site metal ions may be a consideration in the design of inhibitors that selectively target either the mononuclear or binuclear enzymes.

The mononuclear metal center of type-I dihydroorotase from aquifex aeolicus.,Edwards BF, Fernando R, Martin PD, Grimley E, Cordes M, Vaishnav A, Brunzelle JS, Evans HG, Evans DR BMC Biochem. 2013 Dec 9;14(1):36. PMID:24314009^[1]

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

References

↑ Edwards BF, Fernando R, Martin PD, Grimley E, Cordes M, Vaishnav A, Brunzelle JS, Evans HG, Evans DR. The mononuclear metal center of type-I dihydroorotase from aquifex aeolicus. BMC Biochem. 2013 Dec 9;14(1):36. PMID:24314009 doi:http://dx.doi.org/10.1186/1471-2091-14-36

1 Structural highlights
2 Publication Abstract from PubMed
3 References

Proteopedia Page Contributors and Editors (what is this?)

OCA

Retrieved from "http://52.214.119.220/wiki/index.php/4bjh"

@@ Line 1: / Line 1: @@
 ==Crystal Structure of the Aquifex Reactor Complex Formed by Dihydroorotase (H180A, H232A) with Dihydroorotate and Aspartate Transcarbamoylase with N-(phosphonacetyl)-L-aspartate (PALA)==
 <StructureSection load='4bjh' size='340' side='right' caption='[[4bjh]], [[Resolution|resolution]] 2.20&Aring;' scene=''>
@@ Line 5: / Line 6: @@
 </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=BA:BARIUM+ION'>BA</scene>, <scene name='pdbligand=DOR:(4S)-2,6-DIOXOHEXAHYDROPYRIMIDINE-4-CARBOXYLIC+ACID'>DOR</scene>, <scene name='pdbligand=EDO:1,2-ETHANEDIOL'>EDO</scene>, <scene name='pdbligand=PAL:N-(PHOSPHONACETYL)-L-ASPARTIC+ACID'>PAL</scene>, <scene name='pdbligand=PO4:PHOSPHATE+ION'>PO4</scene>, <scene name='pdbligand=ZN:ZINC+ION'>ZN</scene></td></tr>
 <tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/Dihydroorotase Dihydroorotase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.5.2.3 3.5.2.3] </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=4bjh FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4bjh OCA], [http://www.rcsb.org/pdb/explore.do?structureId=4bjh RCSB], [http://www.ebi.ac.uk/pdbsum/4bjh PDBsum]</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=4bjh FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4bjh OCA], [http://pdbe.org/4bjh PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=4bjh RCSB], [http://www.ebi.ac.uk/pdbsum/4bjh PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=4bjh ProSAT]</span></td></tr>
 </table>
 <div style="background-color:#fffaf0;">
@@ Line 15: / Line 16: @@
 From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
 </div>
+<div class="pdbe-citations 4bjh" style="background-color:#fffaf0;"></div>
 == References ==
 <references/>

4bjh

From Proteopedia

Revision as of 11:43, 4 August 2016

Crystal Structure of the Aquifex Reactor Complex Formed by Dihydroorotase (H180A, H232A) with Dihydroorotate and Aspartate Transcarbamoylase with N-(phosphonacetyl)-L-aspartate (PALA)

Structural highlights

Publication Abstract from PubMed

References

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