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| | <StructureSection load='3puh' size='340' side='right'caption='[[3puh]], [[Resolution|resolution]] 2.30Å' scene=''> | | <StructureSection load='3puh' size='340' side='right'caption='[[3puh]], [[Resolution|resolution]] 2.30Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[3puh]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Rhosm Rhosm]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=3PUH OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=3PUH FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[3puh]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Rhodococcus_sp._MB1_'Bresler_1999' Rhodococcus sp. MB1 'Bresler 1999']. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=3PUH OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=3PUH FirstGlance]. <br> |
| - | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=GOL:GLYCEROL'>GOL</scene>, <scene name='pdbligand=SO4:SULFATE+ION'>SO4</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]] 2.3Å</td></tr> |
| - | <tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat"><div style='overflow: auto; max-height: 3em;'>[[3i2f|3i2f]], [[3i2g|3i2g]], [[3i2h|3i2h]], [[3i2i|3i2i]], [[3i2j|3i2j]], [[1ju3|1ju3]], [[3ida|3ida]], [[1ju4|1ju4]], [[3pui|3pui]]</div></td></tr> | + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=GOL:GLYCEROL'>GOL</scene>, <scene name='pdbligand=SO4:SULFATE+ION'>SO4</scene></td></tr> |
| - | <tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">cocE ([https://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=104109 RHOSM])</td></tr>
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| | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=3puh FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=3puh OCA], [https://pdbe.org/3puh PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=3puh RCSB], [https://www.ebi.ac.uk/pdbsum/3puh PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=3puh 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=3puh FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=3puh OCA], [https://pdbe.org/3puh PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=3puh RCSB], [https://www.ebi.ac.uk/pdbsum/3puh PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=3puh ProSAT]</span></td></tr> |
| | </table> | | </table> |
| | == Function == | | == Function == |
| - | [[https://www.uniprot.org/uniprot/COCE_RHOSM COCE_RHOSM]] Hydrolyzes cocaine to benzoate and ecgonine methyl ester, endowing the bacteria with the ability to utilize cocaine as a sole source of carbon and energy for growth, as this bacterium lives in the rhizosphere of coca plants. Also efficiently hydrolyzes cocaethylene, a more potent cocaine metabolite that has been observed in patients who concurrently abuse cocaine and alcohol. Is able to prevent cocaine-induced convulsions and lethality in rat.<ref>PMID:10698749</ref> <ref>PMID:16968810</ref> <ref>PMID:12369817</ref>
| + | [https://www.uniprot.org/uniprot/COCE_RHOSM COCE_RHOSM] Hydrolyzes cocaine to benzoate and ecgonine methyl ester, endowing the bacteria with the ability to utilize cocaine as a sole source of carbon and energy for growth, as this bacterium lives in the rhizosphere of coca plants. Also efficiently hydrolyzes cocaethylene, a more potent cocaine metabolite that has been observed in patients who concurrently abuse cocaine and alcohol. Is able to prevent cocaine-induced convulsions and lethality in rat.<ref>PMID:10698749</ref> <ref>PMID:16968810</ref> <ref>PMID:12369817</ref> |
| - | <div style="background-color:#fffaf0;">
| + | |
| - | == Publication Abstract from PubMed ==
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| - | No small molecule therapeutic is available to treat cocaine addiction, but enzyme-based therapy to accelerate cocaine hydrolysis systemically has recently gained momentum. Bacterial cocaine esterase (CocE) is the fastest known native enzyme that hydrolyzes cocaine. However, its lability at 37 degrees C has limited its therapeutic potential. Crosslinking subunits through disulfide bridging is commonly used to stabilize multimeric enzymes. Herein we utilize structural methods to guide the introduction of two cysteine residues within dimer interface of CocE to facilitate intermolecular disulfide bond formation. The disulphide-crosslinked enzyme displays improved thermostability, particularly when combined with previously described mutations that enhance stability (T172R-G173Q). The newly modified enzyme yielded an extremely stable form of CocE (CCRQ-CocE) that retained greater than 90% of its activity after 41 days at 37 degrees C , representing greater than a 4700-fold improvement over the wild-type enzyme. CCRQ-CocE could also be modified by polyethylene glycol (PEG) polymers, which improved its in vivo residence time from 24 to 72 hours, as measured by a cocaine lethality assay, by self-administration in rodents, and by measurement of inhibition of cocaine-induced cardiovascular effects in Rhesus monkeys. PEG-CCRQ elicited negligible immune response in rodents. Subunit stabilization and PEGylation has thus produced a potential protein therapeutic with markedly higher stability both in vitro and in vivo.
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| - | Subunit Stabilization and PEGylation of Cocaine Esterase Improves In Vivo Residence Time.,Narasimhan D, Collins GT, Nance MR, Nichols J, Edwald E, Chan J, Ko MC, Woods JH, Tesmer JJ, Sunahara RK Mol Pharmacol. 2011 Sep 2. PMID:21890748<ref>PMID:21890748</ref>
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| - | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br>
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| - | </div>
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| - | <div class="pdbe-citations 3puh" style="background-color:#fffaf0;"></div>
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| | ==See Also== | | ==See Also== |
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| | </StructureSection> | | </StructureSection> |
| | [[Category: Large Structures]] | | [[Category: Large Structures]] |
| - | [[Category: Rhosm]] | + | [[Category: Rhodococcus sp. MB1 'Bresler 1999']] |
| - | [[Category: Nance, M R]] | + | [[Category: Nance MR]] |
| - | [[Category: Tesmer, J J.G]] | + | [[Category: Tesmer JJG]] |
| - | [[Category: Alpha/beta hydrolase]]
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| - | [[Category: Cleavage of cocaine]]
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| - | [[Category: Hydrolase]]
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| - | [[Category: Jelly-roll beta-barrel]]
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