PCHB_PSEAE
ID PCHB_PSEAE Reviewed; 101 AA.
AC Q51507;
DT 01-NOV-1997, integrated into UniProtKB/Swiss-Prot.
DT 08-DEC-2000, sequence version 2.
DT 03-AUG-2022, entry version 131.
DE RecName: Full=Isochorismate pyruvate lyase {ECO:0000303|PubMed:11937513};
DE Short=IPL {ECO:0000303|PubMed:11937513};
DE EC=4.2.99.21 {ECO:0000269|PubMed:11937513, ECO:0000269|PubMed:16248620, ECO:0000269|PubMed:19432488, ECO:0000269|PubMed:21751784};
DE AltName: Full=Chorismate mutase {ECO:0000303|PubMed:11937513};
DE Short=CM {ECO:0000303|PubMed:11937513};
DE EC=5.4.99.5 {ECO:0000269|PubMed:11937513, ECO:0000269|PubMed:19432488, ECO:0000269|PubMed:21751784};
DE AltName: Full=Salicylate biosynthesis protein {ECO:0000305|PubMed:7500944};
GN Name=pchB {ECO:0000303|PubMed:7500944}; OrderedLocusNames=PA4230;
OS Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM
OS 14847 / LMG 12228 / 1C / PRS 101 / PAO1).
OC Bacteria; Proteobacteria; Gammaproteobacteria; Pseudomonadales;
OC Pseudomonadaceae; Pseudomonas.
OX NCBI_TaxID=208964;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, PATHWAY, AND INDUCTION.
RC STRAIN=ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C /
RC PRS 101 / PAO1;
RX PubMed=7500944; DOI=10.1007/bf00290369;
RA Serino L., Reimmann C., Baur H., Beyeler M., Visca P., Haas D.;
RT "Structural genes for salicylate biosynthesis from chorismate in
RT Pseudomonas aeruginosa.";
RL Mol. Gen. Genet. 249:217-228(1995).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C /
RC PRS 101 / PAO1;
RX PubMed=10984043; DOI=10.1038/35023079;
RA Stover C.K., Pham X.-Q.T., Erwin A.L., Mizoguchi S.D., Warrener P.,
RA Hickey M.J., Brinkman F.S.L., Hufnagle W.O., Kowalik D.J., Lagrou M.,
RA Garber R.L., Goltry L., Tolentino E., Westbrock-Wadman S., Yuan Y.,
RA Brody L.L., Coulter S.N., Folger K.R., Kas A., Larbig K., Lim R.M.,
RA Smith K.A., Spencer D.H., Wong G.K.-S., Wu Z., Paulsen I.T., Reizer J.,
RA Saier M.H. Jr., Hancock R.E.W., Lory S., Olson M.V.;
RT "Complete genome sequence of Pseudomonas aeruginosa PAO1, an opportunistic
RT pathogen.";
RL Nature 406:959-964(2000).
RN [3]
RP PROTEIN SEQUENCE OF 1-10, FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL
RP PROPERTIES, MUTAGENESIS OF ILE-87, ACTIVITY REGULATION, AND SUBUNIT.
RX PubMed=11937513; DOI=10.1074/jbc.m202410200;
RA Gaille C., Kast P., Haas D.;
RT "Salicylate biosynthesis in Pseudomonas aeruginosa. Purification and
RT characterization of PchB, a novel bifunctional enzyme displaying
RT isochorismate pyruvate-lyase and chorismate mutase activities.";
RL J. Biol. Chem. 277:21768-21775(2002).
RN [4]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND REACTION
RP MECHANISM.
RX PubMed=16248620; DOI=10.1021/ja055871t;
RA DeClue M.S., Baldridge K.K., Kuenzler D.E., Kast P., Hilvert D.;
RT "Isochorismate pyruvate lyase: a pericyclic reaction mechanism?";
RL J. Am. Chem. Soc. 127:15002-15003(2005).
RN [5]
RP MUTAGENESIS OF ALA-37.
RX PubMed=19835359; DOI=10.1021/ja905271g;
RA Marti S., Andres J., Moliner V., Silla E., Tunon I., Bertran J.;
RT "Mechanism and plasticity of isochorismate pyruvate lyase: a computational
RT study.";
RL J. Am. Chem. Soc. 131:16156-16161(2009).
RN [6]
RP X-RAY CRYSTALLOGRAPHY (1.95 ANGSTROMS) IN COMPLEX WITH SUBSTRATE, FUNCTION,
RP SUBUNIT, AND REACTION MECHANISM.
RX PubMed=16914555; DOI=10.1074/jbc.m605470200;
RA Zaitseva J., Lu J., Olechoski K.L., Lamb A.L.;
RT "Two crystal structures of the isochorismate pyruvate lyase from
RT Pseudomonas aeruginosa.";
RL J. Biol. Chem. 281:33441-33449(2006).
RN [7]
RP X-RAY CRYSTALLOGRAPHY (2.25 ANGSTROMS) OF 1-98 OF MUTANTS ALA-42 AND THR-87
RP IN COMPLEX WITH SUBSTRATE, FUNCTION, CATALYTIC ACTIVITY, MUTAGENESIS OF
RP LYS-42 AND ILE-87, BIOPHYSICOCHEMICAL PROPERTIES, AND SUBUNIT.
RX PubMed=19432488; DOI=10.1021/bi900456e;
RA Luo Q., Olucha J., Lamb A.L.;
RT "Structure-function analyses of isochorismate-pyruvate lyase from
RT Pseudomonas aeruginosa suggest differing catalytic mechanisms for the two
RT pericyclic reactions of this bifunctional enzyme.";
RL Biochemistry 48:5239-5245(2009).
RN [8]
RP X-RAY CRYSTALLOGRAPHY (1.79 ANGSTROMS) OF WILD-TYPE AND MUTANT HIS-42 IN
RP COMPLEX WITH SUBSTRATE, FUNCTION, CATALYTIC ACTIVITY, MUTAGENESIS OF
RP LYS-42, AND SUBUNIT.
RX PubMed=21751784; DOI=10.1021/bi200599j;
RA Olucha J., Ouellette A.N., Luo Q., Lamb A.L.;
RT "pH Dependence of catalysis by Pseudomonas aeruginosa isochorismate-
RT pyruvate lyase: implications for transition state stabilization and the
RT role of lysine 42.";
RL Biochemistry 50:7198-7207(2011).
CC -!- FUNCTION: Involved in the incorporation of salicylate into the
CC siderophore pyochelin. Catalyzes the elimination of the enolpyruvyl
CC side chain from isochorismate to yield salicylate and pyruvate via a
CC rare pericyclic hydrogen transfer mechanism from C2 to C5. PchB also
CC catalyzes the nonphysiological Claisen rearrangement of chorismate to
CC prephenate in which the pyruvylenol tail is transferred from a C3 ether
CC linkage to a C1-C9 linkage. {ECO:0000269|PubMed:11937513,
CC ECO:0000269|PubMed:16248620, ECO:0000269|PubMed:16914555,
CC ECO:0000269|PubMed:19432488, ECO:0000269|PubMed:21751784,
CC ECO:0000269|PubMed:7500944}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=isochorismate = pyruvate + salicylate; Xref=Rhea:RHEA:27874,
CC ChEBI:CHEBI:15361, ChEBI:CHEBI:29780, ChEBI:CHEBI:30762;
CC EC=4.2.99.21; Evidence={ECO:0000269|PubMed:11937513,
CC ECO:0000269|PubMed:16248620, ECO:0000269|PubMed:19432488,
CC ECO:0000269|PubMed:21751784};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=chorismate = prephenate; Xref=Rhea:RHEA:13897,
CC ChEBI:CHEBI:29748, ChEBI:CHEBI:29934; EC=5.4.99.5;
CC Evidence={ECO:0000269|PubMed:11937513, ECO:0000269|PubMed:19432488,
CC ECO:0000269|PubMed:21751784};
CC -!- ACTIVITY REGULATION: Inhibited by endo-oxabicyclic diacid resembling to
CC the conformation of the transition state.
CC {ECO:0000269|PubMed:11937513}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=1.05 uM for isochorismate (at pH 7.5 and 30 degrees Celsius)
CC {ECO:0000269|PubMed:16248620};
CC KM=4.3 uM for isochorismate (at pH 7.5 and 22 degrees Celsius)
CC {ECO:0000269|PubMed:19432488};
CC KM=12.5 uM for isochorismate (at pH 7 and 30 degrees Celsius)
CC {ECO:0000269|PubMed:11937513};
CC KM=120 uM for chorismate (at pH 7.5 and 22 degrees Celsius)
CC {ECO:0000269|PubMed:19432488};
CC KM=150 uM for chorismate (at pH 7 and 30 degrees Celsius)
CC {ECO:0000269|PubMed:11937513};
CC Vmax=0.049 umol/min/mg enzyme for isochorismate pyruvate lyase
CC activity (at pH 7 and 30 degrees Celsius)
CC {ECO:0000269|PubMed:11937513};
CC Vmax=0.034 umol/min/mg enzyme for chorismate mutase activity (at pH 7
CC and 30 degrees Celsius) {ECO:0000269|PubMed:11937513};
CC Note=kcat is 1.01 sec(-1) for lyase activity (at pH 7.5 and 22
CC degrees Celsius). kcat is 23.5 sec(-1) for chorismate mutase activity
CC (at pH 7.5 and 22 degrees Celsius). kcat is 106 min(-1) for lyase
CC activity (at pH 7 and 30 degrees Celsius). kcat is 177 sec(-1) for
CC lyase activity (at pH 7.5 and 22 degrees Celsius). kcat is 78 min(-1)
CC for mutase activity (at pH 7 and 30 degrees Celsius). kcat is 106
CC min(-1) for lyase activity (at pH 7 and 30 degrees Celsius).
CC {ECO:0000269|PubMed:11937513, ECO:0000269|PubMed:16248620,
CC ECO:0000269|PubMed:19432488};
CC pH dependence:
CC Optimum pH is 6.8. {ECO:0000269|PubMed:11937513};
CC -!- PATHWAY: Siderophore biosynthesis; salicylate biosynthesis.
CC {ECO:0000269|PubMed:7500944}.
CC -!- SUBUNIT: Dimer of dimers. {ECO:0000269|PubMed:11937513,
CC ECO:0000269|PubMed:16914555, ECO:0000269|PubMed:19432488,
CC ECO:0000269|PubMed:21751784}.
CC -!- INDUCTION: Repressed by iron. {ECO:0000305|PubMed:7500944}.
CC -!- SEQUENCE CAUTION:
CC Sequence=CAA57968.1; Type=Erroneous initiation; Note=Extended N-terminus.; Evidence={ECO:0000305};
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DR EMBL; X82644; CAA57968.1; ALT_INIT; Genomic_DNA.
DR EMBL; AE004091; AAG07618.1; -; Genomic_DNA.
DR PIR; A83117; A83117.
DR PIR; S60202; S58228.
DR RefSeq; NP_252920.1; NC_002516.2.
DR RefSeq; WP_003106950.1; NZ_QZGE01000028.1.
DR PDB; 2H9C; X-ray; 2.35 A; A/B=1-99.
DR PDB; 2H9D; X-ray; 1.95 A; A/B/C/D=1-101.
DR PDB; 3HGW; X-ray; 2.25 A; A/B/C/D=1-98.
DR PDB; 3HGX; X-ray; 2.50 A; A/B=1-99.
DR PDB; 3REM; X-ray; 1.95 A; A/B=1-101.
DR PDB; 3RET; X-ray; 1.79 A; A/B=1-101.
DR PDBsum; 2H9C; -.
DR PDBsum; 2H9D; -.
DR PDBsum; 3HGW; -.
DR PDBsum; 3HGX; -.
DR PDBsum; 3REM; -.
DR PDBsum; 3RET; -.
DR AlphaFoldDB; Q51507; -.
DR SMR; Q51507; -.
DR STRING; 287.DR97_3681; -.
DR PaxDb; Q51507; -.
DR PRIDE; Q51507; -.
DR EnsemblBacteria; AAG07618; AAG07618; PA4230.
DR GeneID; 881846; -.
DR KEGG; pae:PA4230; -.
DR PATRIC; fig|208964.12.peg.4431; -.
DR PseudoCAP; PA4230; -.
DR HOGENOM; CLU_131518_2_1_6; -.
DR InParanoid; Q51507; -.
DR OMA; LIHWFIN; -.
DR PhylomeDB; Q51507; -.
DR BioCyc; MetaCyc:MON-15305; -.
DR BioCyc; PAER208964:G1FZ6-4303-MON; -.
DR BRENDA; 4.2.99.21; 5087.
DR SABIO-RK; Q51507; -.
DR UniPathway; UPA00025; -.
DR EvolutionaryTrace; Q51507; -.
DR PHI-base; PHI:5058; -.
DR Proteomes; UP000002438; Chromosome.
DR GO; GO:0016835; F:carbon-oxygen lyase activity; IDA:UniProtKB.
DR GO; GO:0004106; F:chorismate mutase activity; IDA:UniProtKB.
DR GO; GO:0043904; F:isochorismate pyruvate lyase activity; IEA:UniProtKB-EC.
DR GO; GO:0046417; P:chorismate metabolic process; IBA:GO_Central.
DR GO; GO:0042864; P:pyochelin biosynthetic process; IDA:UniProtKB.
DR GO; GO:0009697; P:salicylic acid biosynthetic process; IDA:PseudoCAP.
DR Gene3D; 1.20.59.10; -; 1.
DR InterPro; IPR036263; Chorismate_II_sf.
DR InterPro; IPR036979; CM_dom_sf.
DR InterPro; IPR002701; CM_II_prokaryot.
DR InterPro; IPR008241; Isochorismate_pyruvate-lyase.
DR Pfam; PF01817; CM_2; 1.
DR PIRSF; PIRSF029775; Isochor_pyr_lyas; 1.
DR SMART; SM00830; CM_2; 1.
DR SUPFAM; SSF48600; SSF48600; 1.
DR TIGRFAMs; TIGR01803; CM-like; 1.
DR PROSITE; PS51168; CHORISMATE_MUT_2; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Direct protein sequencing; Isomerase; Lyase;
KW Reference proteome.
FT CHAIN 1..101
FT /note="Isochorismate pyruvate lyase"
FT /id="PRO_0000119198"
FT DOMAIN 4..94
FT /note="Chorismate mutase"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00515"
FT BINDING 14
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:16914555,
FT ECO:0000269|PubMed:21751784"
FT BINDING 31
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:16914555,
FT ECO:0000269|PubMed:19432488, ECO:0000269|PubMed:21751784"
FT BINDING 42
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:16914555,
FT ECO:0000269|PubMed:21751784"
FT BINDING 90
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:16914555,
FT ECO:0000269|PubMed:19432488, ECO:0000269|PubMed:21751784"
FT MUTAGEN 37
FT /note="A->I: Increases the rate constant for the mutase
FT activity by a factor of 1000, and also increases the lyase
FT catalytic efficiency by a factor of 6."
FT /evidence="ECO:0000269|PubMed:19835359"
FT MUTAGEN 42
FT /note="K->A: Active across the entire pH range from 4 to 9.
FT 11-fold reduction of the affinity for isochorismate and 7-
FT fold reduction of the catalytic efficiency for lyase
FT activity. 6-fold reduction of the affinity for chorismate
FT and 15-fold reduction of the catalytic efficiency for
FT mutase activity."
FT /evidence="ECO:0000269|PubMed:19432488,
FT ECO:0000269|PubMed:21751784"
FT MUTAGEN 42
FT /note="K->E: Loss of both lyase and mutase activity at any
FT pH tested."
FT /evidence="ECO:0000269|PubMed:19432488,
FT ECO:0000269|PubMed:21751784"
FT MUTAGEN 42
FT /note="K->H: At pH 5, 15-fold reduction of the affinity for
FT isochorismate, but only a slight reduction of the catalytic
FT efficiency for lyase activity. At pH 7.5, 13-fold reduction
FT of the affinity for isochorismate and 4-fold reduction of
FT the catalytic efficiency for lyase activity. At pH 5,
FT strong reduction of the affinity for chorismate, but only a
FT 2-fold reduction of the catalytic efficiency for mutase
FT activity. At pH 7.5, strong reduction of the affinity for
FT chorismate, but only a slight reduction of the catalytic
FT efficiency for mutase activity."
FT /evidence="ECO:0000269|PubMed:19432488,
FT ECO:0000269|PubMed:21751784"
FT MUTAGEN 42
FT /note="K->Q: Loss of mutase activity. 15-fold reduction of
FT the affinity for isochorismate and 3-fold reduction of the
FT catalytic efficiency for isochorismate-pyruvate lyase
FT activity."
FT /evidence="ECO:0000269|PubMed:19432488"
FT MUTAGEN 43
FT /note="A->P: Slight reduction of the affinity for
FT isochorismate and of the catalytic efficiency for
FT isochorismate-pyruvate lyase activity. Slight reduction of
FT the affinity for chorismate and of the catalytic efficiency
FT for mutase activity."
FT /evidence="ECO:0000269|PubMed:19432488"
FT MUTAGEN 87
FT /note="I->T: 4-fold reduction of the affinity for
FT isochorismate and 3-fold reduction of the catalytic
FT efficiency for isochorismate-pyruvate lyase activity. 4-
FT fold reduction of the affinity for chorismate and 15-fold
FT reduction of the catalytic efficiency for mutase activity."
FT /evidence="ECO:0000269|PubMed:11937513,
FT ECO:0000269|PubMed:19432488"
FT HELIX 4..6
FT /evidence="ECO:0007829|PDB:3RET"
FT HELIX 10..38
FT /evidence="ECO:0007829|PDB:3RET"
FT HELIX 39..41
FT /evidence="ECO:0007829|PDB:3RET"
FT HELIX 45..47
FT /evidence="ECO:0007829|PDB:3RET"
FT HELIX 51..67
FT /evidence="ECO:0007829|PDB:3RET"
FT HELIX 72..96
FT /evidence="ECO:0007829|PDB:3RET"
SQ SEQUENCE 101 AA; 11432 MW; 97AE6DCE7A6ABD7D CRC64;
MKTPEDCTGL ADIREAIDRI DLDIVQALGR RMDYVKAASR FKASEAAIPA PERVAAMLPE
RARWAEENGL DAPFVEGLFA QIIHWYIAEQ IKYWRQTRGA A
