COCE_RHOSM
ID COCE_RHOSM Reviewed; 574 AA.
AC Q9L9D7;
DT 31-JAN-2002, integrated into UniProtKB/Swiss-Prot.
DT 01-OCT-2000, sequence version 1.
DT 03-AUG-2022, entry version 117.
DE RecName: Full=Cocaine esterase;
DE EC=3.1.1.84;
GN Name=cocE;
OS Rhodococcus sp. (strain MB1 Bresler).
OC Bacteria; Actinobacteria; Corynebacteriales; Nocardiaceae; Rhodococcus.
OX NCBI_TaxID=104109;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 1-11, FUNCTION,
RP CATALYTIC ACTIVITY, BIOTECHNOLOGY, AND INDUCTION.
RC STRAIN=MB1;
RX PubMed=10698749; DOI=10.1128/aem.66.3.904-908.2000;
RA Bresler M.M., Rosser S.J., Basran A., Bruce N.C.;
RT "Gene cloning and nucleotide sequencing and properties of a cocaine
RT esterase from Rhodococcus sp. strain MB1.";
RL Appl. Environ. Microbiol. 66:904-908(2000).
RN [2]
RP FUNCTION, TEMPERATURE DEPENDENCE, AND MUTAGENESIS OF TYR-44 AND SER-117.
RC STRAIN=MB1;
RX PubMed=16968810; DOI=10.1124/mol.106.025999;
RA Cooper Z.D., Narasimhan D., Sunahara R.K., Mierzejewski P.,
RA Jutkiewicz E.M., Larsen N.A., Wilson I.A., Landry D.W., Woods J.H.;
RT "Rapid and robust protection against cocaine-induced lethality in rats by
RT the bacterial cocaine esterase.";
RL Mol. Pharmacol. 70:1885-1891(2006).
RN [3]
RP X-RAY CRYSTALLOGRAPHY (1.58 ANGSTROMS) IN COMPLEXES WITH BENZOATE AND
RP TRANSITION STATE ANALOG, ACTIVE SITE, REACTION MECHANISM, AND DOMAIN.
RC STRAIN=MB1;
RX PubMed=11742345; DOI=10.1038/nsb742;
RA Larsen N.A., Turner J.M., Stevens J., Rosser S.J., Basran A., Lerner R.A.,
RA Bruce N.C., Wilson I.A.;
RT "Crystal structure of a bacterial cocaine esterase.";
RL Nat. Struct. Biol. 9:17-21(2002).
RN [4]
RP X-RAY CRYSTALLOGRAPHY (1.6 ANGSTROMS) OF MUTANTS PHE-44 AND ALA-117 IN
RP COMPLEX WITH BENZOATE, FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL
RP PROPERTIES, BIOTECHNOLOGY, AND MUTAGENESIS OF TYR-44; GLN-55; SER-117;
RP TRP-151; TRP-166; ASP-259; PHE-261; HIS-287; LEU-407 AND PHE-408.
RC STRAIN=MB1;
RX PubMed=12369817; DOI=10.1021/bi026131p;
RA Turner J.M., Larsen N.A., Basran A., Barbas C.F. III, Bruce N.C.,
RA Wilson I.A., Lerner R.A.;
RT "Biochemical characterization and structural analysis of a highly
RT proficient cocaine esterase.";
RL Biochemistry 41:12297-12307(2002).
RN [5]
RP X-RAY CRYSTALLOGRAPHY (1.51 ANGSTROMS) OF WILD-TYPE AND MUTANTS LYS-169;
RP ARG-172; GLN-173 AND ARG-172/GLN-173, MUTAGENESIS OF LEU-169; THR-172 AND
RP GLY-173, CATALYTIC ACTIVITY, AND SUBUNIT.
RC STRAIN=MB1;
RX PubMed=20436035; DOI=10.1093/protein/gzq025;
RA Narasimhan D., Nance M.R., Gao D., Ko M.C., Macdonald J., Tamburi P.,
RA Yoon D., Landry D.M., Woods J.H., Zhan C.G., Tesmer J.J., Sunahara R.K.;
RT "Structural analysis of thermostabilizing mutations of cocaine esterase.";
RL Protein Eng. Des. Sel. 23:537-547(2010).
RN [6]
RP X-RAY CRYSTALLOGRAPHY (1.53 ANGSTROMS) OF WILD-TYPE AND A
RP DISULFIDE-STABILIZED DIMERIC MUTANT.
RC STRAIN=MB1;
RX PubMed=21890748; DOI=10.1124/mol.111.074997;
RA Narasimhan D., Collins G.T., Nance M.R., Nichols J., Edwald E., Chan J.,
RA Ko M.C., Woods J.H., Tesmer J.J., Sunahara R.K.;
RT "Subunit stabilization and polyethylene glycolation of cocaine esterase
RT improves in vivo residence time.";
RL Mol. Pharmacol. 80:1056-1065(2011).
CC -!- FUNCTION: Hydrolyzes cocaine to benzoate and ecgonine methyl ester,
CC endowing the bacteria with the ability to utilize cocaine as a sole
CC source of carbon and energy for growth, as this bacterium lives in the
CC rhizosphere of coca plants. Also efficiently hydrolyzes cocaethylene, a
CC more potent cocaine metabolite that has been observed in patients who
CC concurrently abuse cocaine and alcohol. Is able to prevent cocaine-
CC induced convulsions and lethality in rat. {ECO:0000269|PubMed:10698749,
CC ECO:0000269|PubMed:12369817, ECO:0000269|PubMed:16968810}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=cocaine + H2O = benzoate + ecgonine methyl ester + H(+);
CC Xref=Rhea:RHEA:27506, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:16150, ChEBI:CHEBI:59908, ChEBI:CHEBI:60056; EC=3.1.1.84;
CC Evidence={ECO:0000269|PubMed:10698749, ECO:0000269|PubMed:12369817,
CC ECO:0000269|PubMed:20436035};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.64 uM for cocaine {ECO:0000269|PubMed:12369817};
CC KM=1.6 uM for cocaethylene {ECO:0000269|PubMed:12369817};
CC Note=kcat is 7.8 sec(-1) with cocaine as substrate, and 9.4 sec(-1)
CC with cocaethylene.;
CC pH dependence:
CC Optimum pH is 9.0. {ECO:0000269|PubMed:12369817};
CC Temperature dependence:
CC Is relatively unstable at physiological temperatures since it
CC displays a half-life of 13 minutes in rat plasma at 37 degrees
CC Celsius. {ECO:0000269|PubMed:12369817, ECO:0000269|PubMed:16968810};
CC -!- PATHWAY: Alkaloid degradation; cocaine degradation.
CC -!- SUBUNIT: Homodimer. The protein aggregates upon heat inactivation.
CC {ECO:0000269|PubMed:12369817, ECO:0000269|PubMed:20436035}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000305}.
CC -!- INDUCTION: Positively induced by cocaine.
CC {ECO:0000269|PubMed:10698749}.
CC -!- DOMAIN: It consists of three domains: domain 1 contains the active
CC site; domains 2 and 3 are involved in substrate recognition. Domain 1
CC contains the GxSxxG motif found in most members of the alpha/beta
CC hydrolase superfamily. {ECO:0000269|PubMed:11742345}.
CC -!- BIOTECHNOLOGY: Because of the high catalytic proficiency of CocE, it is
CC an attractive candidate for novel protein-based therapies for cocaine
CC overdose, as this cocaine-degrading enzyme could be used for rapid
CC cocaine detoxification in an emergency setting. However, wild-type CocE
CC is relatively unstable, but this can be improved by specific mutations.
CC Thus, improved stability of engineered CocE enzymes will have a
CC profound influence on the use of this protein to combat cocaine-induced
CC toxicity and addiction in humans. Has also a potential as a highly-
CC sensitive drug detector. {ECO:0000269|PubMed:10698749,
CC ECO:0000269|PubMed:12369817}.
CC -!- MISCELLANEOUS: This enzyme hydrolyzes cocaine faster than any other
CC known cocaine esterase.
CC -!- MISCELLANEOUS: Incorporating disulfide bonds between cysteine residues
CC substituted at Gly-4 and Ser-10 conveys significant improvements to the
CC thermostability and the half-life at 37 degrees Celsius. Moreover, in
CC combination with T172R/G173Q mutations, the disulfide-stabilized dimer
CC (CCRQ-CocE) remains more than 90% active for longer than 40 days at 37
CC degrees Celsius, representing a >4700-fold improvement over wt-CocE.
CC The enhanced stability serves as a better substrate for modification,
CC with polyethylene glycol (PEG) moieties providing the therapeutic with
CC stealth properties. PEGylated CCRQ-CocE retains full in vitro enzymatic
CC activity, protects rodents up to 72 hours in a cocaine overdose model,
CC diminishes self-administration for 72 hours in rats, reduces cocaine-
CC induced cardiovascular effects and locomotor functions in monkeys for
CC up to 48 hours, and displays reduced immunogenicity in mice.
CC -!- SIMILARITY: Belongs to the CocE/NonD hydrolase family. {ECO:0000305}.
CC -!- WEB RESOURCE: Name=Protein Spotlight; Note=Rhodococcus: Nature's junkie
CC - Issue 27 of October 2002;
CC URL="https://web.expasy.org/spotlight/back_issues/027";
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DR EMBL; AF173165; AAF42807.1; -; Genomic_DNA.
DR PDB; 1JU3; X-ray; 1.58 A; A=1-574.
DR PDB; 1JU4; X-ray; 1.63 A; A=1-574.
DR PDB; 1L7Q; X-ray; 1.76 A; A=1-574.
DR PDB; 1L7R; X-ray; 1.64 A; A=1-574.
DR PDB; 3I2F; X-ray; 2.50 A; A=1-574.
DR PDB; 3I2G; X-ray; 2.50 A; A=1-574.
DR PDB; 3I2H; X-ray; 1.65 A; A=1-574.
DR PDB; 3I2I; X-ray; 2.14 A; A=1-574.
DR PDB; 3I2J; X-ray; 2.01 A; A=1-574.
DR PDB; 3I2K; X-ray; 1.51 A; A=1-574.
DR PDB; 3IDA; X-ray; 1.60 A; A=1-574.
DR PDB; 3PUH; X-ray; 2.30 A; A/B=1-574.
DR PDB; 3PUI; X-ray; 1.53 A; A=1-574.
DR PDB; 4P08; X-ray; 2.34 A; A=4-574.
DR PDB; 7F65; X-ray; 2.20 A; A=1-574.
DR PDBsum; 1JU3; -.
DR PDBsum; 1JU4; -.
DR PDBsum; 1L7Q; -.
DR PDBsum; 1L7R; -.
DR PDBsum; 3I2F; -.
DR PDBsum; 3I2G; -.
DR PDBsum; 3I2H; -.
DR PDBsum; 3I2I; -.
DR PDBsum; 3I2J; -.
DR PDBsum; 3I2K; -.
DR PDBsum; 3IDA; -.
DR PDBsum; 3PUH; -.
DR PDBsum; 3PUI; -.
DR PDBsum; 4P08; -.
DR PDBsum; 7F65; -.
DR AlphaFoldDB; Q9L9D7; -.
DR SMR; Q9L9D7; -.
DR DrugBank; DB03793; Benzoic acid.
DR DrugBank; DB01795; Phenylboronic acid.
DR ESTHER; rhosm-cocE; Cocaine_esterase.
DR KEGG; ag:AAF42807; -.
DR BioCyc; MetaCyc:MON-15371; -.
DR BRENDA; 3.1.1.84; 5397.
DR UniPathway; UPA00110; -.
DR EvolutionaryTrace; Q9L9D7; -.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0052689; F:carboxylic ester hydrolase activity; IEA:UniProtKB-KW.
DR GO; GO:0008239; F:dipeptidyl-peptidase activity; IEA:InterPro.
DR GO; GO:0050784; P:cocaine catabolic process; IEA:UniProtKB-UniPathway.
DR Gene3D; 3.40.50.1820; -; 1.
DR InterPro; IPR029058; AB_hydrolase.
DR InterPro; IPR005674; CocE/Ser_esterase.
DR InterPro; IPR008979; Galactose-bd-like_sf.
DR InterPro; IPR000383; Xaa-Pro-like_dom.
DR InterPro; IPR013736; Xaa-Pro_dipept_C.
DR Pfam; PF02129; Peptidase_S15; 1.
DR Pfam; PF08530; PepX_C; 1.
DR SMART; SM00939; PepX_C; 1.
DR SUPFAM; SSF49785; SSF49785; 1.
DR SUPFAM; SSF53474; SSF53474; 1.
DR TIGRFAMs; TIGR00976; NonD; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Cytoplasm; Direct protein sequencing; Hydrolase;
KW Serine esterase.
FT CHAIN 1..574
FT /note="Cocaine esterase"
FT /id="PRO_0000090000"
FT REGION 1..144
FT /note="1A"
FT REGION 145..240
FT /note="2"
FT REGION 241..354
FT /note="1B"
FT REGION 355..574
FT /note="3"
FT ACT_SITE 117
FT /note="Acyl-ester intermediate"
FT /evidence="ECO:0000269|PubMed:11742345"
FT ACT_SITE 259
FT /note="Charge relay system"
FT /evidence="ECO:0000269|PubMed:11742345"
FT ACT_SITE 287
FT /note="Charge relay system"
FT /evidence="ECO:0000269|PubMed:11742345"
FT BINDING 44
FT /ligand="substrate"
FT BINDING 118
FT /ligand="substrate"
FT SITE 44
FT /note="Probably involved in activating the substrate
FT carbonyl and the acyl enzyme for hydrolysis"
FT MUTAGEN 44
FT /note="Y->F: Loss of activity. Has no protective effects
FT against cocaine-induced convulsions and lethality in rat."
FT /evidence="ECO:0000269|PubMed:12369817,
FT ECO:0000269|PubMed:16968810"
FT MUTAGEN 55
FT /note="Q->A,E: Decrease in activity."
FT /evidence="ECO:0000269|PubMed:12369817"
FT MUTAGEN 117
FT /note="S->A: Loss of activity. Has no protective effects
FT against cocaine-induced convulsions and lethality in rat."
FT /evidence="ECO:0000269|PubMed:12369817,
FT ECO:0000269|PubMed:16968810"
FT MUTAGEN 117
FT /note="S->C: Great decrease in activity."
FT /evidence="ECO:0000269|PubMed:12369817,
FT ECO:0000269|PubMed:16968810"
FT MUTAGEN 151
FT /note="W->A: Decrease in activity."
FT /evidence="ECO:0000269|PubMed:12369817"
FT MUTAGEN 166
FT /note="W->A: Decrease in activity."
FT /evidence="ECO:0000269|PubMed:12369817"
FT MUTAGEN 169
FT /note="L->K: Displays greatly enhanced stability, with a
FT half-life of 570 minutes at 37 degrees Celsius. Exhibits
FT 4.5-fold reduction in catalytic efficiency."
FT /evidence="ECO:0000269|PubMed:20436035"
FT MUTAGEN 172
FT /note="T->R: Displays enhanced stability, with a half-life
FT of 78 minutes at 37 degrees Celsius, and exhibits 3-fold
FT reduction in catalytic efficiency. Displays enhanced
FT stability, with a half-life of 370 minutes at 37 degrees
FT Celsius, and exhibits 3-fold reduction in catalytic
FT efficiency; when associated with Q-173."
FT /evidence="ECO:0000269|PubMed:20436035"
FT MUTAGEN 173
FT /note="G->Q: Displays enhanced stability, with a half-life
FT of 75 minutes at 37 degrees Celsius, and has no deleterious
FT effect on catalytic efficiency. Displays enhanced
FT stability, with a half-life of 370 minutes at 37 degrees
FT Celsius, and exhibits 3-fold reduction in catalytic
FT efficiency; when associated with R-172."
FT /evidence="ECO:0000269|PubMed:20436035"
FT MUTAGEN 259
FT /note="D->N: Loss of activity."
FT /evidence="ECO:0000269|PubMed:12369817"
FT MUTAGEN 261
FT /note="F->A: Decrease in activity."
FT /evidence="ECO:0000269|PubMed:12369817"
FT MUTAGEN 287
FT /note="H->A: Loss of activity."
FT /evidence="ECO:0000269|PubMed:12369817"
FT MUTAGEN 407
FT /note="L->A: Decrease in activity."
FT /evidence="ECO:0000269|PubMed:12369817"
FT MUTAGEN 408
FT /note="F->A: Decrease in activity."
FT /evidence="ECO:0000269|PubMed:12369817"
FT STRAND 6..15
FT /evidence="ECO:0007829|PDB:3I2K"
FT STRAND 21..29
FT /evidence="ECO:0007829|PDB:3I2K"
FT STRAND 35..44
FT /evidence="ECO:0007829|PDB:3I2K"
FT HELIX 49..53
FT /evidence="ECO:0007829|PDB:3I2K"
FT TURN 54..56
FT /evidence="ECO:0007829|PDB:3I2K"
FT HELIX 60..64
FT /evidence="ECO:0007829|PDB:3I2K"
FT STRAND 68..73
FT /evidence="ECO:0007829|PDB:3I2K"
FT TURN 86..89
FT /evidence="ECO:0007829|PDB:3I2K"
FT HELIX 90..103
FT /evidence="ECO:0007829|PDB:3I2K"
FT STRAND 107..113
FT /evidence="ECO:0007829|PDB:3I2K"
FT HELIX 118..127
FT /evidence="ECO:0007829|PDB:3I2K"
FT STRAND 134..137
FT /evidence="ECO:0007829|PDB:3I2K"
FT STRAND 139..141
FT /evidence="ECO:0007829|PDB:3I2K"
FT HELIX 147..151
FT /evidence="ECO:0007829|PDB:3I2K"
FT HELIX 160..177
FT /evidence="ECO:0007829|PDB:3I2K"
FT STRAND 178..180
FT /evidence="ECO:0007829|PDB:3I2K"
FT HELIX 185..196
FT /evidence="ECO:0007829|PDB:3I2K"
FT HELIX 199..203
FT /evidence="ECO:0007829|PDB:3I2K"
FT STRAND 205..207
FT /evidence="ECO:0007829|PDB:3PUI"
FT HELIX 212..217
FT /evidence="ECO:0007829|PDB:3I2K"
FT HELIX 220..223
FT /evidence="ECO:0007829|PDB:3I2K"
FT TURN 224..227
FT /evidence="ECO:0007829|PDB:3I2K"
FT HELIX 233..236
FT /evidence="ECO:0007829|PDB:3I2K"
FT HELIX 241..244
FT /evidence="ECO:0007829|PDB:3I2K"
FT STRAND 251..258
FT /evidence="ECO:0007829|PDB:3I2K"
FT HELIX 262..272
FT /evidence="ECO:0007829|PDB:3I2K"
FT TURN 273..275
FT /evidence="ECO:0007829|PDB:3I2K"
FT STRAND 278..286
FT /evidence="ECO:0007829|PDB:3I2K"
FT STRAND 291..294
FT /evidence="ECO:0007829|PDB:3I2K"
FT HELIX 301..303
FT /evidence="ECO:0007829|PDB:3I2K"
FT HELIX 307..322
FT /evidence="ECO:0007829|PDB:3I2K"
FT TURN 326..331
FT /evidence="ECO:0007829|PDB:3I2K"
FT STRAND 334..340
FT /evidence="ECO:0007829|PDB:3I2K"
FT TURN 341..343
FT /evidence="ECO:0007829|PDB:3I2K"
FT STRAND 344..352
FT /evidence="ECO:0007829|PDB:3I2K"
FT STRAND 357..364
FT /evidence="ECO:0007829|PDB:3I2K"
FT STRAND 377..381
FT /evidence="ECO:0007829|PDB:3I2K"
FT STRAND 387..393
FT /evidence="ECO:0007829|PDB:3I2K"
FT STRAND 407..410
FT /evidence="ECO:0007829|PDB:3I2K"
FT HELIX 419..421
FT /evidence="ECO:0007829|PDB:3I2K"
FT STRAND 428..431
FT /evidence="ECO:0007829|PDB:3I2K"
FT STRAND 439..457
FT /evidence="ECO:0007829|PDB:3I2K"
FT STRAND 459..467
FT /evidence="ECO:0007829|PDB:3I2K"
FT STRAND 473..482
FT /evidence="ECO:0007829|PDB:3I2K"
FT HELIX 483..485
FT /evidence="ECO:0007829|PDB:3I2K"
FT STRAND 489..491
FT /evidence="ECO:0007829|PDB:3I2K"
FT STRAND 501..514
FT /evidence="ECO:0007829|PDB:3I2K"
FT STRAND 519..526
FT /evidence="ECO:0007829|PDB:3I2K"
FT STRAND 537..540
FT /evidence="ECO:0007829|PDB:3I2K"
FT HELIX 542..544
FT /evidence="ECO:0007829|PDB:3I2K"
FT HELIX 547..549
FT /evidence="ECO:0007829|PDB:3I2K"
FT STRAND 553..563
FT /evidence="ECO:0007829|PDB:3I2K"
FT STRAND 566..572
FT /evidence="ECO:0007829|PDB:3I2K"
SQ SEQUENCE 574 AA; 62132 MW; 9E35724F586089B7 CRC64;
MVDGNYSVAS NVMVPMRDGV RLAVDLYRPD ADGPVPVLLV RNPYDKFDVF AWSTQSTNWL
EFVRDGYAVV IQDTRGLFAS EGEFVPHVDD EADAEDTLSW ILEQAWCDGN VGMFGVSYLG
VTQWQAAVSG VGGLKAIAPS MASADLYRAP WYGPGGALSV EALLGWSALI GTGLITSRSD
ARPEDAADFV QLAAILNDVA GAASVTPLAE QPLLGRLIPW VIDQVVDHPD NDESWQSISL
FERLGGLATP ALITAGWYDG FVGESLRTFV AVKDNADARL VVGPWSHSNL TGRNADRKFG
IAATYPIQEA TTMHKAFFDR HLRGETDALA GVPKVRLFVM GIDEWRDETD WPLPDTAYTP
FYLGGSGAAN TSTGGGTLST SISGTESADT YLYDPADPVP SLGGTLLFHN GDNGPADQRP
IHDRDDVLCY STEVLTDPVE VTGTVSARLF VSSSAVDTDF TAKLVDVFPD GRAIALCDGI
VRMRYRETLV NPTLIEAGEI YEVAIDMLAT SNVFLPGHRI MVQVSSSNFP KYDRNSNTGG
VIAREQLEEM CTAVNRIHRG PEHPSHIVLP IIKR
