HAD_PSEUY
ID HAD_PSEUY Reviewed; 232 AA.
AC Q53464;
DT 15-JUL-1998, integrated into UniProtKB/Swiss-Prot.
DT 01-NOV-1996, sequence version 1.
DT 03-AUG-2022, entry version 89.
DE RecName: Full=(S)-2-haloacid dehalogenase {ECO:0000305};
DE EC=3.8.1.2 {ECO:0000269|PubMed:7490277, ECO:0000269|PubMed:9614112};
DE AltName: Full=2-haloalkanoic acid dehalogenase;
DE AltName: Full=Halocarboxylic acid halidohydrolase;
DE AltName: Full=L-2-haloacid dehalogenase {ECO:0000303|PubMed:7944368};
DE AltName: Full=L-DEX;
OS Pseudomonas sp. (strain YL).
OC Bacteria; Proteobacteria.
OX NCBI_TaxID=66693;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RX PubMed=7944368; DOI=10.1128/aem.60.9.3375-3380.1994;
RA Nardi-Dei V., Kurihara T., Okamura T., Liu J.Q., Koshikawa H., Ozaki H.,
RA Terashima Y., Esaki N., Soda K.;
RT "Comparative studies of genes encoding thermostable L-2-halo acid
RT dehalogenase from Pseudomonas sp. strain YL, other dehalogenases, and two
RT related hypothetical proteins from Escherichia coli.";
RL Appl. Environ. Microbiol. 60:3375-3380(1994).
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, ACTIVE SITE,
RP AND MUTAGENESIS OF ASP-10; THR-14; ARG-41; SER-118; LYS-151; TYR-157;
RP SER-175; ASN-177 AND ASP-180.
RX PubMed=7490277; DOI=10.1093/oxfordjournals.jbchem.a124861;
RA Kurihara T., Liu J.-Q., Nardi-Dei V., Koshikawa H., Esaki N., Soda K.;
RT "Comprehensive site-directed mutagenesis of L-2-halo acid dehalogenase to
RT probe catalytic amino acid residues.";
RL J. Biochem. 117:1317-1322(1995).
RN [3] {ECO:0007744|PDB:1JUD}
RP X-RAY CRYSTALLOGRAPHY (2.5 ANGSTROMS), AND SUBUNIT.
RX PubMed=8702766; DOI=10.1074/jbc.271.34.20322;
RA Hisano T., Hata Y., Fujii T., Liu J.-Q., Kurihara T., Esaki N., Soda K.;
RT "Crystal structure of L-2-haloacid dehalogenase from Pseudomonas sp. YL. An
RT alpha/beta hydrolase structure that is different from the alpha/beta
RT hydrolase fold.";
RL J. Biol. Chem. 271:20322-20330(1996).
RN [4] {ECO:0007744|PDB:1ZRM, ECO:0007744|PDB:1ZRN}
RP X-RAY CRYSTALLOGRAPHY (1.83 ANGSTROMS) OF MUTANT ALA-175 IN COMPLEX WITH
RP MONOCHLOROACETATE AND 2-CHLORO-N-BUTYRATE, FUNCTION, CATALYTIC ACTIVITY,
RP ACTIVE SITE, AND MUTAGENESIS OF SER-175.
RX PubMed=9614112; DOI=10.1074/jbc.273.24.15035;
RA Li Y.F., Hata Y., Fujii T., Hisano T., Nishihara M., Kurihara T., Esaki N.;
RT "Crystal structures of reaction intermediates of L-2-haloacid dehalogenase
RT and implications for the reaction mechanism.";
RL J. Biol. Chem. 273:15035-15044(1998).
RN [5] {ECO:0007744|PDB:1QH9}
RP X-RAY CRYSTALLOGRAPHY (2.50 ANGSTROMS) IN COMPLEX WITH LACTATE.
RA Li Y.-F., Hata Y., Fujii T., Kurihara T., Esaki N.;
RT "The Structure of L-2-Haloacid Dehalogenase Complexed with a Reaction
RT Product Reveals the Mechanism of Intermediate Hydrolysis in Dehalogenase.";
RL Submitted (MAY-1999) to the PDB data bank.
CC -!- FUNCTION: Catalyzes the hydrolytic dehalogenation of small (S)-2-
CC haloalkanoic acids to yield the corresponding (R)-2-hydroxyalkanoic
CC acids (PubMed:7490277, PubMed:9614112). Acts on acids of short chain
CC lengths, C(2) to C(4), with inversion of configuration at C-2
CC (PubMed:7490277, PubMed:9614112). Active with 2-halogenated carboxylic
CC acids and converts only the S-isomer (or L-isomer) of 2-chloropropionic
CC acid with inversion of configuration to produce R-lactate (or D-isomer)
CC (PubMed:7490277, PubMed:9614112). {ECO:0000269|PubMed:7490277,
CC ECO:0000269|PubMed:9614112}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=an (S)-2-haloacid + H2O = a (2R)-2-hydroxycarboxylate + a
CC halide anion + H(+); Xref=Rhea:RHEA:11192, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:16042, ChEBI:CHEBI:58314,
CC ChEBI:CHEBI:137405; EC=3.8.1.2; Evidence={ECO:0000269|PubMed:7490277,
CC ECO:0000269|PubMed:9614112};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(S)-2-chloropropanoate + H2O = (R)-lactate + chloride + H(+);
CC Xref=Rhea:RHEA:67956, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:16004, ChEBI:CHEBI:17996, ChEBI:CHEBI:73934;
CC Evidence={ECO:0000269|PubMed:7490277, ECO:0000269|PubMed:9614112};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.37 mM for (2S)-2-chloropropanoic acid
CC {ECO:0000269|PubMed:7490277};
CC Note=kcat is 47 sec(-1) for (2S)-2-chloropropanoic acid as substrate.
CC {ECO:0000269|PubMed:7490277};
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:8702766}.
CC -!- BIOTECHNOLOGY: (S)-2-haloacid dehalogenases may be used for the
CC biodegradation of halogenated substances and their derivatives which
CC are widely used as pesticides, herbicides and other industrial
CC products. {ECO:0000305}.
CC -!- SIMILARITY: Belongs to the HAD-like hydrolase superfamily. S-2-
CC haloalkanoic acid dehalogenase family. {ECO:0000305}.
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DR EMBL; S74078; AAB32245.1; -; Genomic_DNA.
DR PDB; 1JUD; X-ray; 2.50 A; A=1-232.
DR PDB; 1QH9; X-ray; 2.50 A; A=1-232.
DR PDB; 1ZRM; X-ray; 2.00 A; A=1-232.
DR PDB; 1ZRN; X-ray; 1.83 A; A=1-232.
DR PDBsum; 1JUD; -.
DR PDBsum; 1QH9; -.
DR PDBsum; 1ZRM; -.
DR PDBsum; 1ZRN; -.
DR AlphaFoldDB; Q53464; -.
DR SMR; Q53464; -.
DR DrugBank; DB03568; Butyric Acid.
DR DrugBank; DB03066; D-Lactic acid.
DR PRIDE; Q53464; -.
DR KEGG; ag:AAB32245; -.
DR EvolutionaryTrace; Q53464; -.
DR GO; GO:0018784; F:(S)-2-haloacid dehalogenase activity; IEA:UniProtKB-EC.
DR CDD; cd02588; HAD_L2-DEX; 1.
DR Gene3D; 1.10.150.240; -; 1.
DR Gene3D; 3.40.50.1000; -; 1.
DR InterPro; IPR006328; 2-HAD.
DR InterPro; IPR036412; HAD-like_sf.
DR InterPro; IPR006439; HAD-SF_hydro_IA.
DR InterPro; IPR041492; HAD_2.
DR InterPro; IPR023214; HAD_sf.
DR InterPro; IPR023198; PGP-like_dom2.
DR Pfam; PF13419; HAD_2; 1.
DR PRINTS; PR00413; HADHALOGNASE.
DR SUPFAM; SSF56784; SSF56784; 1.
DR TIGRFAMs; TIGR01493; HAD-SF-IA-v2; 1.
DR TIGRFAMs; TIGR01428; HAD_type_II; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Hydrolase.
FT CHAIN 1..232
FT /note="(S)-2-haloacid dehalogenase"
FT /id="PRO_0000079166"
FT REGION 175..180
FT /note="Important for catalytic activity"
FT /evidence="ECO:0000269|PubMed:7490277,
FT ECO:0000305|PubMed:9614112"
FT ACT_SITE 10
FT /note="Nucleophile"
FT /evidence="ECO:0000269|PubMed:9614112,
FT ECO:0000305|PubMed:7490277"
FT BINDING 11..12
FT /ligand="an (S)-2-haloacid"
FT /ligand_id="ChEBI:CHEBI:137405"
FT /evidence="ECO:0000269|PubMed:9614112, ECO:0000269|Ref.5,
FT ECO:0007744|PDB:1QH9, ECO:0007744|PDB:1ZRM,
FT ECO:0007744|PDB:1ZRN"
FT BINDING 41
FT /ligand="an (S)-2-haloacid"
FT /ligand_id="ChEBI:CHEBI:137405"
FT /evidence="ECO:0000305|PubMed:9614112,
FT ECO:0007744|PDB:1ZRM, ECO:0007744|PDB:1ZRN"
FT BINDING 118..119
FT /ligand="an (S)-2-haloacid"
FT /ligand_id="ChEBI:CHEBI:137405"
FT /evidence="ECO:0000269|PubMed:9614112, ECO:0000269|Ref.5,
FT ECO:0007744|PDB:1QH9, ECO:0007744|PDB:1ZRM,
FT ECO:0007744|PDB:1ZRN"
FT SITE 14
FT /note="Important for catalytic activity"
FT /evidence="ECO:0000269|PubMed:7490277,
FT ECO:0000305|PubMed:9614112"
FT SITE 151
FT /note="Important for catalytic activity"
FT /evidence="ECO:0000269|PubMed:7490277,
FT ECO:0000305|PubMed:9614112"
FT SITE 157
FT /note="Important for catalytic activity"
FT /evidence="ECO:0000269|PubMed:7490277,
FT ECO:0000305|PubMed:9614112"
FT MUTAGEN 10
FT /note="D->E,S,A,G: Loss of catalytic activity."
FT /evidence="ECO:0000269|PubMed:7490277"
FT MUTAGEN 10
FT /note="D->N: Severe reduction in catalytic activity."
FT /evidence="ECO:0000269|PubMed:7490277"
FT MUTAGEN 14
FT /note="T->A: Severe reduction in catalytic activity. No
FT effect on susbtrate affinity."
FT /evidence="ECO:0000269|PubMed:7490277"
FT MUTAGEN 14
FT /note="T->P,C: Severe reduction in catalytic activity."
FT /evidence="ECO:0000269|PubMed:7490277"
FT MUTAGEN 14
FT /note="T->S: Mild reduction in catalytic activity."
FT /evidence="ECO:0000269|PubMed:7490277"
FT MUTAGEN 41
FT /note="R->E: Severe reduction in catalytic activity."
FT /evidence="ECO:0000269|PubMed:7490277"
FT MUTAGEN 41
FT /note="R->K: Severe reduction in catalytic activity. 4-fold
FT decrease in substrate affinity."
FT /evidence="ECO:0000269|PubMed:7490277"
FT MUTAGEN 41
FT /note="R->W: Loss of catalytic activity."
FT /evidence="ECO:0000269|PubMed:7490277"
FT MUTAGEN 118
FT /note="S->A: Severe reduction in catalytic activity. 25-
FT fold decrease in substrate affinity."
FT /evidence="ECO:0000269|PubMed:7490277"
FT MUTAGEN 151
FT /note="K->A: Loss of catalytic activity."
FT /evidence="ECO:0000269|PubMed:7490277"
FT MUTAGEN 151
FT /note="K->R: Severe reduction in catalytic activity."
FT /evidence="ECO:0000269|PubMed:7490277"
FT MUTAGEN 157
FT /note="Y->C,S,H,W: Severe reduction in catalytic activity."
FT /evidence="ECO:0000269|PubMed:7490277"
FT MUTAGEN 157
FT /note="Y->F: Severe reduction in catalytic activity. 1.5-
FT fold decrease in substrate affinity."
FT /evidence="ECO:0000269|PubMed:7490277"
FT MUTAGEN 175
FT /note="S->A: Severe reduction in catalytic activity. 2-fold
FT decrease in substrate affinity."
FT /evidence="ECO:0000269|PubMed:7490277,
FT ECO:0000269|PubMed:9614112"
FT MUTAGEN 175
FT /note="S->C: Severe reduction in catalytic activity."
FT /evidence="ECO:0000269|PubMed:7490277"
FT MUTAGEN 175
FT /note="S->T: Moderate reduction in catalytic activity."
FT /evidence="ECO:0000269|PubMed:7490277"
FT MUTAGEN 177
FT /note="N->D,I: Loss of catalytic activity."
FT /evidence="ECO:0000269|PubMed:7490277"
FT MUTAGEN 177
FT /note="N->W,K,H,Q: Severe reduction in catalytic activity."
FT /evidence="ECO:0000269|PubMed:7490277"
FT MUTAGEN 180
FT /note="D->N,E,S,G: Loss of catalytic activity."
FT /evidence="ECO:0000269|PubMed:7490277"
FT STRAND 6..9
FT /evidence="ECO:0007829|PDB:1ZRN"
FT TURN 13..15
FT /evidence="ECO:0007829|PDB:1ZRN"
FT HELIX 19..28
FT /evidence="ECO:0007829|PDB:1ZRN"
FT TURN 30..32
FT /evidence="ECO:0007829|PDB:1ZRN"
FT HELIX 33..54
FT /evidence="ECO:0007829|PDB:1ZRN"
FT HELIX 60..75
FT /evidence="ECO:0007829|PDB:1ZRN"
FT HELIX 81..89
FT /evidence="ECO:0007829|PDB:1ZRN"
FT HELIX 90..93
FT /evidence="ECO:0007829|PDB:1ZRN"
FT HELIX 100..109
FT /evidence="ECO:0007829|PDB:1ZRN"
FT STRAND 113..120
FT /evidence="ECO:0007829|PDB:1ZRN"
FT HELIX 122..131
FT /evidence="ECO:0007829|PDB:1ZRN"
FT HELIX 135..137
FT /evidence="ECO:0007829|PDB:1ZRN"
FT STRAND 139..144
FT /evidence="ECO:0007829|PDB:1ZRN"
FT HELIX 145..147
FT /evidence="ECO:0007829|PDB:1ZRN"
FT HELIX 154..164
FT /evidence="ECO:0007829|PDB:1ZRN"
FT HELIX 168..170
FT /evidence="ECO:0007829|PDB:1ZRN"
FT STRAND 171..176
FT /evidence="ECO:0007829|PDB:1ZRN"
FT HELIX 178..187
FT /evidence="ECO:0007829|PDB:1ZRN"
FT STRAND 191..194
FT /evidence="ECO:0007829|PDB:1ZRN"
FT STRAND 203..205
FT /evidence="ECO:0007829|PDB:1ZRN"
FT STRAND 209..214
FT /evidence="ECO:0007829|PDB:1ZRN"
FT HELIX 215..219
FT /evidence="ECO:0007829|PDB:1ZRN"
SQ SEQUENCE 232 AA; 26177 MW; D276B5164B02E2C1 CRC64;
MDYIKGIAFD LYGTLFDVHS VVGRCDEAFP GRGREISALW RQKQLEYTWL RSLMNRYVNF
QQATEDALRF TCRHLGLDLD ARTRSTLCDA YLRLAPFSEV PDSLRELKRR GLKLAILSNG
SPQSIDAVVS HAGLRDGFDH LLSVDPVQVY KPDNRVYELA EQALGLDRSA ILFVSSNAWD
ATGARYFGFP TCWINRTGNV FEEMGQTPDW EVTSLRAVVE LFETAAGKAE KG
