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| | ==Cocaine Esterase with mutations G4C, S10C== | | ==Cocaine Esterase with mutations G4C, S10C== |
| - | <StructureSection load='3pui' size='340' side='right' caption='[[3pui]], [[Resolution|resolution]] 1.53Å' scene=''> | + | <StructureSection load='3pui' size='340' side='right'caption='[[3pui]], [[Resolution|resolution]] 1.53Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[3pui]] is a 1 chain structure with sequence from [http://en.wikipedia.org/wiki/Rhosm Rhosm]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=3PUI OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=3PUI FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[3pui]] is a 1 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=3PUI OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=3PUI FirstGlance]. <br> |
| - | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=CL:CHLORIDE+ION'>CL</scene>, <scene name='pdbligand=NA:SODIUM+ION'>NA</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]] 1.53Å</td></tr> |
| - | <tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[3i2f|3i2f]], [[3i2g|3i2g]], [[3i2h|3i2h]], [[3i2i|3i2i]], [[3i2j|3i2j]], [[1ju3|1ju3]], [[1ju4|1ju4]], [[3ida|3ida]], [[3puh|3puh]]</td></tr> | + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=CL:CHLORIDE+ION'>CL</scene>, <scene name='pdbligand=NA:SODIUM+ION'>NA</scene></td></tr> |
| - | <tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">cocE ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=104109 RHOSM])</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=3pui FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=3pui OCA], [https://pdbe.org/3pui PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=3pui RCSB], [https://www.ebi.ac.uk/pdbsum/3pui PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=3pui 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=3pui FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=3pui OCA], [http://pdbe.org/3pui PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=3pui RCSB], [http://www.ebi.ac.uk/pdbsum/3pui PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=3pui ProSAT]</span></td></tr> | + | |
| | </table> | | </table> |
| | == Function == | | == Function == |
| - | [[http://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;"> | | <div style="background-color:#fffaf0;"> |
| | == Publication Abstract from PubMed == | | == Publication Abstract from PubMed == |
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| | __TOC__ | | __TOC__ |
| | </StructureSection> | | </StructureSection> |
| - | [[Category: Rhosm]] | + | [[Category: Large Structures]] |
| - | [[Category: Nance, M R]] | + | [[Category: Rhodococcus sp. MB1 'Bresler 1999']] |
| - | [[Category: Tesmer, J J.G]]
| + | [[Category: Nance MR]] |
| - | [[Category: Alpha/beta hydrolase]] | + | [[Category: Tesmer JJG]] |
| - | [[Category: Cleavage of cocaine]] | + | |
| - | [[Category: Hydrolase]]
| + | |
| - | [[Category: Jelly-roll beta-barrel]]
| + | |
| Structural highlights
Function
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.[1] [2] [3]
Publication Abstract from PubMed
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.
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[4]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
See Also
References
- ↑ Bresler MM, Rosser SJ, Basran A, Bruce NC. Gene cloning and nucleotide sequencing and properties of a cocaine esterase from Rhodococcus sp. strain MB1. Appl Environ Microbiol. 2000 Mar;66(3):904-8. PMID:10698749
- ↑ Cooper ZD, Narasimhan D, Sunahara RK, Mierzejewski P, Jutkiewicz EM, Larsen NA, Wilson IA, Landry DW, Woods JH. Rapid and robust protection against cocaine-induced lethality in rats by the bacterial cocaine esterase. Mol Pharmacol. 2006 Dec;70(6):1885-91. Epub 2006 Sep 12. PMID:16968810 doi:10.1124/mol.106.025999
- ↑ Turner JM, Larsen NA, Basran A, Barbas CF 3rd, Bruce NC, Wilson IA, Lerner RA. Biochemical characterization and structural analysis of a highly proficient cocaine esterase. Biochemistry. 2002 Oct 15;41(41):12297-307. PMID:12369817
- ↑ Narasimhan D, Collins GT, Nance MR, Nichols J, Edwald E, Chan J, Ko MC, Woods JH, Tesmer JJ, Sunahara RK. Subunit Stabilization and PEGylation of Cocaine Esterase Improves In Vivo Residence Time. Mol Pharmacol. 2011 Sep 2. PMID:21890748 doi:10.1124/mol.111.074997
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