SubmitCancel

Skip Header

You are using a version of browser that may not display all the features of this website. Please consider upgrading your browser.

Q9L9D7

- COCE_RHOSM

UniProt

Q9L9D7 - COCE_RHOSM

(max 400 entries)x

Your basket is currently empty.

Select item(s) and click on "Add to basket" to create your own collection here
(400 entries max)

Protein
Cocaine esterase
Gene
cocE
Organism
Rhodococcus sp. (strain MB1 Bresler)
Status
Reviewed - Annotation score: 5 out of 5 - Experimental evidence at protein leveli

Functioni

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.3 Publications

Catalytic activityi

Cocaine + H2O = ecgonine methyl ester + benzoate.3 Publications

Kineticsi

kcat is 7.8 sec(-1) with cocaine as substrate, and 9.4 sec(-1) with cocaethylene.

  1. KM=0.64 µM for cocaine1 Publication
  2. KM=1.6 µM for cocaethylene

pH dependencei

Optimum pH is 9.0.

Temperature dependencei

Is relatively unstable at physiological temperatures since it displays a half-life of 13 minutes in rat plasma at 37 degrees Celsius.

Pathwayi

Sites

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Binding sitei44 – 441Substrate
Sitei44 – 441Probably involved in activating the substrate carbonyl and the acyl enzyme for hydrolysis
Active sitei117 – 1171Acyl-ester intermediate1 Publication
Binding sitei118 – 1181Substrate; via amide nitrogen
Active sitei259 – 2591Charge relay system1 Publication
Active sitei287 – 2871Charge relay system1 Publication

GO - Molecular functioni

  1. aminopeptidase activity Source: InterPro
  2. dipeptidyl-peptidase activity Source: InterPro

GO - Biological processi

  1. cocaine catabolic process Source: UniProtKB-UniPathway
Complete GO annotation...

Keywords - Molecular functioni

Hydrolase, Serine esterase

Enzyme and pathway databases

BioCyciMetaCyc:MONOMER-15371.
BRENDAi3.1.1.1. 5397.
UniPathwayiUPA00110.

Names & Taxonomyi

Protein namesi
Recommended name:
Cocaine esterase (EC:3.1.1.84)
Gene namesi
Name:cocE
OrganismiRhodococcus sp. (strain MB1 Bresler)
Taxonomic identifieri104109 [NCBI]
Taxonomic lineageiBacteriaActinobacteriaActinobacteridaeActinomycetalesCorynebacterineaeNocardiaceaeRhodococcus

Subcellular locationi

Cytoplasm Inferred

GO - Cellular componenti

  1. cytoplasm Source: UniProtKB-SubCell
Complete GO annotation...

Keywords - Cellular componenti

Cytoplasm

Pathology & Biotechi

Biotechnological usei

Because of the high catalytic proficiency of CocE, it is an attractive candidate for novel protein-based therapies for cocaine overdose, as this cocaine-degrading enzyme could be used for rapid cocaine detoxification in an emergency setting. However, wild-type CocE is relatively unstable, but this can be improved by specific mutations. Thus, improved stability of engineered CocE enzymes will have a profound influence on the use of this protein to combat cocaine-induced toxicity and addiction in humans. Has also a potential as a highly-sensitive drug detector.2 Publications

Mutagenesis

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Mutagenesisi44 – 441Y → F: Loss of activity. Has no protective effects against cocaine-induced convulsions and lethality in rat. 2 Publications
Mutagenesisi55 – 551Q → A or E: Decrease in activity. 1 Publication
Mutagenesisi117 – 1171S → A: Loss of activity. Has no protective effects against cocaine-induced convulsions and lethality in rat. 2 Publications
Mutagenesisi117 – 1171S → C: Great decrease in activity. 2 Publications
Mutagenesisi151 – 1511W → A: Decrease in activity. 1 Publication
Mutagenesisi166 – 1661W → A: Decrease in activity. 1 Publication
Mutagenesisi169 – 1691L → K: Displays greatly enhanced stability, with a half-life of 570 minutes at 37 degrees Celsius. Exhibits 4.5-fold reduction in catalytic efficiency. 1 Publication
Mutagenesisi172 – 1721T → R: Displays enhanced stability, with a half-life of 78 minutes at 37 degrees Celsius, and exhibits 3-fold reduction in catalytic efficiency. Displays enhanced stability, with a half-life of 370 minutes at 37 degrees Celsius, and exhibits 3-fold reduction in catalytic efficiency; when associated with Q-173. 1 Publication
Mutagenesisi173 – 1731G → Q: Displays enhanced stability, with a half-life of 75 minutes at 37 degrees Celsius, and has no deleterious effect on catalytic efficiency. Displays enhanced stability, with a half-life of 370 minutes at 37 degrees Celsius, and exhibits 3-fold reduction in catalytic efficiency; when associated with R-172. 1 Publication
Mutagenesisi259 – 2591D → N: Loss of activity. 1 Publication
Mutagenesisi261 – 2611F → A: Decrease in activity. 1 Publication
Mutagenesisi287 – 2871H → A: Loss of activity. 1 Publication
Mutagenesisi407 – 4071L → A: Decrease in activity. 1 Publication
Mutagenesisi408 – 4081F → A: Decrease in activity. 1 Publication

PTM / Processingi

Molecule processing

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Chaini1 – 574574Cocaine esterase
PRO_0000090000Add
BLAST

Expressioni

Inductioni

Positively induced by cocaine.1 Publication

Interactioni

Subunit structurei

Homodimer. The protein aggregates upon heat inactivation.1 Publication

Structurei

Secondary structure

Legend: HelixTurnBeta strand
Show more details
Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Beta strandi6 – 1510
Beta strandi21 – 299
Beta strandi35 – 4410
Helixi49 – 535
Turni54 – 563
Helixi60 – 645
Beta strandi68 – 736
Turni86 – 894
Helixi90 – 10314
Beta strandi107 – 1137
Helixi118 – 12710
Beta strandi134 – 1374
Beta strandi139 – 1413
Helixi147 – 1515
Helixi160 – 17718
Beta strandi178 – 1803
Helixi185 – 19612
Helixi199 – 2035
Beta strandi205 – 2073
Helixi212 – 2176
Helixi220 – 2234
Turni224 – 2274
Helixi233 – 2364
Helixi241 – 2444
Beta strandi251 – 2588
Helixi262 – 27211
Turni273 – 2753
Beta strandi278 – 2869
Beta strandi291 – 2944
Helixi301 – 3033
Helixi307 – 32216
Turni326 – 3316
Beta strandi334 – 3407
Turni341 – 3433
Beta strandi344 – 3529
Beta strandi357 – 3648
Beta strandi377 – 3815
Beta strandi387 – 3937
Beta strandi407 – 4104
Helixi419 – 4213
Beta strandi428 – 4314
Beta strandi439 – 45719
Beta strandi459 – 4679
Beta strandi473 – 48210
Helixi483 – 4853
Beta strandi489 – 4913
Beta strandi501 – 51414
Beta strandi519 – 5268
Beta strandi537 – 5404
Helixi542 – 5443
Helixi547 – 5493
Beta strandi553 – 56311
Beta strandi566 – 5727

3D structure databases

Select the link destinations:
PDBe
RCSB PDB
PDBj
Links Updated
EntryMethodResolution (Å)ChainPositionsPDBsum
1JU3X-ray1.58A1-574[»]
1JU4X-ray1.63A1-574[»]
1L7QX-ray1.76A1-574[»]
1L7RX-ray1.64A1-574[»]
3I2FX-ray2.50A1-574[»]
3I2GX-ray2.50A1-574[»]
3I2HX-ray1.65A1-574[»]
3I2IX-ray2.14A1-574[»]
3I2JX-ray2.01A1-574[»]
3I2KX-ray1.51A1-574[»]
3IDAX-ray1.60A1-574[»]
3PUHX-ray2.30A/B1-574[»]
3PUIX-ray1.53A1-574[»]
ProteinModelPortaliQ9L9D7.
SMRiQ9L9D7. Positions 5-574.

Miscellaneous databases

EvolutionaryTraceiQ9L9D7.

Family & Domainsi

Region

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Regioni1 – 1441441A
Add
BLAST
Regioni145 – 240962
Add
BLAST
Regioni241 – 3541141B
Add
BLAST
Regioni355 – 5742203
Add
BLAST

Domaini

It consists of three domains: domain 1 contains the active site; domains 2 and 3 are involved in substrate recognition. Domain 1 contains the GxSxxG motif found in most members of the alpha/beta hydrolase superfamily.1 Publication

Sequence similaritiesi

Family and domain databases

Gene3Di2.60.120.260. 1 hit.
3.40.50.1820. 2 hits.
InterProiIPR029058. AB_hydrolase.
IPR005674. CocE/Ser_esterase.
IPR008979. Galactose-bd-like.
IPR000383. Xaa-Pro-like_dom.
IPR013736. Xaa-Pro_dipept_C.
[Graphical view]
PfamiPF02129. Peptidase_S15. 1 hit.
PF08530. PepX_C. 1 hit.
[Graphical view]
SMARTiSM00939. PepX_C. 1 hit.
[Graphical view]
SUPFAMiSSF49785. SSF49785. 1 hit.
SSF53474. SSF53474. 1 hit.
TIGRFAMsiTIGR00976. /NonD. 1 hit.

Sequencei

Sequence statusi: Complete.

Q9L9D7-1 [UniParc]FASTAAdd to Basket

« Hide

MVDGNYSVAS NVMVPMRDGV RLAVDLYRPD ADGPVPVLLV RNPYDKFDVF    50
AWSTQSTNWL EFVRDGYAVV IQDTRGLFAS EGEFVPHVDD EADAEDTLSW 100
ILEQAWCDGN VGMFGVSYLG VTQWQAAVSG VGGLKAIAPS MASADLYRAP 150
WYGPGGALSV EALLGWSALI GTGLITSRSD ARPEDAADFV QLAAILNDVA 200
GAASVTPLAE QPLLGRLIPW VIDQVVDHPD NDESWQSISL FERLGGLATP 250
ALITAGWYDG FVGESLRTFV AVKDNADARL VVGPWSHSNL TGRNADRKFG 300
IAATYPIQEA TTMHKAFFDR HLRGETDALA GVPKVRLFVM GIDEWRDETD 350
WPLPDTAYTP FYLGGSGAAN TSTGGGTLST SISGTESADT YLYDPADPVP 400
SLGGTLLFHN GDNGPADQRP IHDRDDVLCY STEVLTDPVE VTGTVSARLF 450
VSSSAVDTDF TAKLVDVFPD GRAIALCDGI VRMRYRETLV NPTLIEAGEI 500
YEVAIDMLAT SNVFLPGHRI MVQVSSSNFP KYDRNSNTGG VIAREQLEEM 550
CTAVNRIHRG PEHPSHIVLP IIKR 574
Length:574
Mass (Da):62,132
Last modified:October 1, 2000 - v1
Checksum:i9E35724F586089B7
GO

Sequence databases

Select the link destinations:
EMBL
GenBank
DDBJ
Links Updated
AF173165 Genomic DNA. Translation: AAF42807.1.

Cross-referencesi

Web resourcesi

Protein Spotlight

Rhodococcus: Nature's junkie - Issue 27 of October 2002

Sequence databases

Select the link destinations:
EMBL
GenBank
DDBJ
Links Updated
AF173165 Genomic DNA. Translation: AAF42807.1 .

3D structure databases

Select the link destinations:
PDBe
RCSB PDB
PDBj
Links Updated
Entry Method Resolution (Å) Chain Positions PDBsum
1JU3 X-ray 1.58 A 1-574 [» ]
1JU4 X-ray 1.63 A 1-574 [» ]
1L7Q X-ray 1.76 A 1-574 [» ]
1L7R X-ray 1.64 A 1-574 [» ]
3I2F X-ray 2.50 A 1-574 [» ]
3I2G X-ray 2.50 A 1-574 [» ]
3I2H X-ray 1.65 A 1-574 [» ]
3I2I X-ray 2.14 A 1-574 [» ]
3I2J X-ray 2.01 A 1-574 [» ]
3I2K X-ray 1.51 A 1-574 [» ]
3IDA X-ray 1.60 A 1-574 [» ]
3PUH X-ray 2.30 A/B 1-574 [» ]
3PUI X-ray 1.53 A 1-574 [» ]
ProteinModelPortali Q9L9D7.
SMRi Q9L9D7. Positions 5-574.
ModBasei Search...
MobiDBi Search...

Protocols and materials databases

Structural Biology Knowledgebase Search...

Enzyme and pathway databases

UniPathwayi UPA00110 .
BioCyci MetaCyc:MONOMER-15371.
BRENDAi 3.1.1.1. 5397.

Miscellaneous databases

EvolutionaryTracei Q9L9D7.

Family and domain databases

Gene3Di 2.60.120.260. 1 hit.
3.40.50.1820. 2 hits.
InterProi IPR029058. AB_hydrolase.
IPR005674. CocE/Ser_esterase.
IPR008979. Galactose-bd-like.
IPR000383. Xaa-Pro-like_dom.
IPR013736. Xaa-Pro_dipept_C.
[Graphical view ]
Pfami PF02129. Peptidase_S15. 1 hit.
PF08530. PepX_C. 1 hit.
[Graphical view ]
SMARTi SM00939. PepX_C. 1 hit.
[Graphical view ]
SUPFAMi SSF49785. SSF49785. 1 hit.
SSF53474. SSF53474. 1 hit.
TIGRFAMsi TIGR00976. /NonD. 1 hit.
ProtoNeti Search...

Publicationsi

  1. "Gene cloning and nucleotide sequencing and properties of a cocaine esterase from Rhodococcus sp. strain MB1."
    Bresler M.M., Rosser S.J., Basran A., Bruce N.C.
    Appl. Environ. Microbiol. 66:904-908(2000) [PubMed] [Europe PMC] [Abstract]
    Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 1-11, FUNCTION, CATALYTIC ACTIVITY, BIOTECHNOLOGY, INDUCTION.
    Strain: MB1.
  2. "Rapid and robust protection against cocaine-induced lethality in rats by the bacterial cocaine esterase."
    Cooper Z.D., Narasimhan D., Sunahara R.K., Mierzejewski P., Jutkiewicz E.M., Larsen N.A., Wilson I.A., Landry D.W., Woods J.H.
    Mol. Pharmacol. 70:1885-1891(2006) [PubMed] [Europe PMC] [Abstract]
    Cited for: FUNCTION, TEMPERATURE DEPENDENCE, MUTAGENESIS OF TYR-44 AND SER-117.
    Strain: MB1.
  3. Cited for: X-RAY CRYSTALLOGRAPHY (1.58 ANGSTROMS) IN COMPLEXES WITH BENZOATE AND TRANSITION STATE ANALOG, ACTIVE SITE, REACTION MECHANISM, DOMAIN.
    Strain: MB1.
  4. "Biochemical characterization and structural analysis of a highly proficient cocaine esterase."
    Turner J.M., Larsen N.A., Basran A., Barbas C.F. III, Bruce N.C., Wilson I.A., Lerner R.A.
    Biochemistry 41:12297-12307(2002) [PubMed] [Europe PMC] [Abstract]
    Cited for: X-RAY CRYSTALLOGRAPHY (1.6 ANGSTROMS) OF MUTANTS PHE-44 AND ALA-117 IN COMPLEX WITH BENZOATE, FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, BIOTECHNOLOGY, MUTAGENESIS OF TYR-44; GLN-55; SER-117; TRP-151; TRP-166; ASP-259; PHE-261; HIS-287; LEU-407 AND PHE-408.
    Strain: MB1.
  5. Cited for: X-RAY CRYSTALLOGRAPHY (1.51 ANGSTROMS) OF WILD-TYPE AND MUTANTS LYS-169; ARG-172; GLN-173 AND ARG-172/GLN-173, MUTAGENESIS OF LEU-169; THR-172 AND GLY-173, CATALYTIC ACTIVITY, SUBUNIT.
    Strain: MB1.
  6. "Subunit stabilization and polyethylene glycolation of cocaine esterase improves in vivo residence time."
    Narasimhan D., Collins G.T., Nance M.R., Nichols J., Edwald E., Chan J., Ko M.C., Woods J.H., Tesmer J.J., Sunahara R.K.
    Mol. Pharmacol. 80:1056-1065(2011) [PubMed] [Europe PMC] [Abstract]
    Cited for: X-RAY CRYSTALLOGRAPHY (1.53 ANGSTROMS) OF WILD-TYPE AND A DISULFIDE-STABILIZED DIMERIC MUTANT.
    Strain: MB1.

Entry informationi

Entry nameiCOCE_RHOSM
AccessioniPrimary (citable) accession number: Q9L9D7
Entry historyi
Integrated into UniProtKB/Swiss-Prot: January 31, 2002
Last sequence update: October 1, 2000
Last modified: September 3, 2014
This is version 85 of the entry and version 1 of the sequence. [Complete history]
Entry statusiReviewed (UniProtKB/Swiss-Prot)
Annotation programProkaryotic Protein Annotation Program

Miscellaneousi

Miscellaneous

This enzyme hydrolyzes cocaine faster than any other known cocaine esterase.
Incorporating disulfide bonds between cysteine residues substituted at Gly-4 and Ser-10 conveys significant improvements to the thermostability and the half-life at 37 degrees Celsius. Moreover, in combination with T172R/G173Q mutations, the disulfide-stabilized dimer (CCRQ-CocE) remains more than 90% active for longer than 40 days at 37 degrees Celsius, representing a >4700-fold improvement over wt-CocE. The enhanced stability serves as a better substrate for modification, with polyethylene glycol (PEG) moieties providing the therapeutic with stealth properties. PEGylated CCRQ-CocE retains full in vitro enzymatic activity, protects rodents up to 72 hours in a cocaine overdose model, diminishes self-administration for 72 hours in rats, reduces cocaine-induced cardiovascular effects and locomotor functions in monkeys for up to 48 hours, and displays reduced immunogenicity in mice.

Keywords - Technical termi

3D-structure, Direct protein sequencing

Documents

  1. PATHWAY comments
    Index of metabolic and biosynthesis pathways
  2. PDB cross-references
    Index of Protein Data Bank (PDB) cross-references
  3. Protein Spotlight
    Protein Spotlight articles and cited UniProtKB/Swiss-Prot entries
  4. SIMILARITY comments
    Index of protein domains and families

External Data

Dasty 3

Similar proteinsi