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4p08

From Proteopedia

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Engineered thermostable dimeric cocaine esterase

Structural highlights

4p08 is a 1 chain structure with sequence from Rhodococcus sp. MB1. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance.
Method:	X-ray diffraction, Resolution 2.341Å
Resources:	FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT

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

Cocaine esterase (CocE) is known as the most efficient natural enzyme for cocaine hydrolysis. The major obstacle to the clinical application of wild-type CocE is the thermoinstability with a half-life of only approximately 12 min at 37 degrees C. The previously designed T172R/G173Q mutant (denoted as enzyme E172-173) with an improved in vitro half-life of approximately 6 h at 37 degrees C is currently in clinical trial Phase II for cocaine overdose treatment. Through molecular modeling and dynamics simulation, we designed and characterized a promising new mutant of E172-173 with extra L196C/I301C mutations (denoted as enzyme E196-301) to produce cross-subunit disulfide bonds that stabilize the dimer structure. The cross-subunit disulfide bonds were confirmed by X-ray diffraction. The designed L196C/I301C mutations have not only considerably extended the in vitro half-life at 37 degrees C to >100 days, but also significantly improved the catalytic efficiency against cocaine by approximately 150%. In addition, the thermostable E196-301 can be PEGylated to significantly prolong the residence time in mice. The PEGylated E196-301 can fully protect mice from a lethal dose of cocaine (180 mg/kg, LD100) for at least 3 days, with an average protection time of approximately 94h. This is the longest in vivo protection of mice from the lethal dose of cocaine demonstrated within all studies using an exogenous enzyme reported so far. Hence, E196-301 may be developed to become a more valuable therapeutic enzyme for cocaine abuse treatment, and it demonstrates that a general design strategy and protocol to simultaneously improve both the stability and function are feasible for rational protein drug design.

Rational Design, Preparation, and Characterization of a Therapeutic Enzyme Mutant with Improved Stability and Function for Cocaine Detoxification.,Fang L, Chow KM, Hou S, Xue L, Chen X, Rodgers DW, Zheng F, Zhan CG ACS Chem Biol. 2014 Jun 11. PMID:24919140^[4]

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

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
↑ Fang L, Chow KM, Hou S, Xue L, Chen X, Rodgers DW, Zheng F, Zhan CG. Rational Design, Preparation, and Characterization of a Therapeutic Enzyme Mutant with Improved Stability and Function for Cocaine Detoxification. ACS Chem Biol. 2014 Jun 11. PMID:24919140 doi:http://dx.doi.org/10.1021/cb500257s

1 Structural highlights
2 Function
3 Publication Abstract from PubMed
4 See Also
5 References

Proteopedia Page Contributors and Editors (what is this?)

OCA

Retrieved from "http://52.214.119.220/wiki/index.php/4p08"

@@ Line 1: / Line 1: @@
 ==Engineered thermostable dimeric cocaine esterase==
-<StructureSection load='4p08' size='340' side='right' caption='[[4p08]], [[Resolution|resolution]] 2.34&Aring;' scene=''>
+<StructureSection load='4p08' size='340' side='right'caption='[[4p08]], [[Resolution|resolution]] 2.34&Aring;' scene=''>
 == Structural highlights ==
-<table><tr><td colspan='2'>[[4p08]] is a 1 chain structure. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4P08 OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4P08 FirstGlance]. <br>
+<table><tr><td colspan='2'>[[4p08]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Rhodococcus_sp._MB1 Rhodococcus sp. MB1]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4P08 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=4P08 FirstGlance]. <br>
-</td></tr><tr><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/Cocaine_esterase Cocaine esterase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.1.1.84 3.1.1.84] </span></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]] 2.341&#8491;</td></tr>
-<tr><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4p08 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4p08 OCA], [http://www.rcsb.org/pdb/explore.do?structureId=4p08 RCSB], [http://www.ebi.ac.uk/pdbsum/4p08 PDBsum]</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=4p08 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4p08 OCA], [https://pdbe.org/4p08 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=4p08 RCSB], [https://www.ebi.ac.uk/pdbsum/4p08 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=4p08 ProSAT]</span></td></tr>
-<table>
+</table>
+== 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>
 <div style="background-color:#fffaf0;">
 == Publication Abstract from PubMed ==
@@ Line 14: / Line 17: @@
 From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
 </div>
+<div class="pdbe-citations 4p08" style="background-color:#fffaf0;"></div>
+==See Also==
+*[[Cocaine esterase|Cocaine esterase]]
 == References ==
 <references/>
 __TOC__
 </StructureSection>
-[[Category: Cocaine esterase]]
+[[Category: Large Structures]]
-[[Category: Chow, K M.]]
+[[Category: Rhodococcus sp. MB1]]
-[[Category: Fang, L.]]
+[[Category: Chow K-M]]
-[[Category: Rodgers, D W.]]
+[[Category: Fang L]]
-[[Category: Zhan, C G.]]
+[[Category: Rodgers DW]]
-[[Category: Cocaine]]
+[[Category: Zhan C-G]]
-[[Category: Disulfide-linked dimer]]
-[[Category: Esterase]]
-[[Category: Hydrolase]]
-[[Category: Mutant]]

4p08

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Current revision

Engineered thermostable dimeric cocaine esterase

Structural highlights

Function

Publication Abstract from PubMed

See Also

References

Contents

Proteopedia Page Contributors and Editors (what is this?)

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