BCPMH_TRICW
ID BCPMH_TRICW Reviewed; 514 AA.
AC Q45087;
DT 20-DEC-2017, integrated into UniProtKB/Swiss-Prot.
DT 01-NOV-1996, sequence version 1.
DT 03-AUG-2022, entry version 85.
DE RecName: Full=Multifunctional alkaline phosphatase superfamily protein PehA {ECO:0000305};
DE AltName: Full=Phosphodiesterase {ECO:0000303|PubMed:8824203};
DE EC=3.1.4.- {ECO:0000269|PubMed:8824203};
DE AltName: Full=Phosphonate ester hydrolase {ECO:0000303|PubMed:8771792, ECO:0000303|PubMed:8824203, ECO:0000312|EMBL:AAC44467.1};
DE Short=PEH {ECO:0000303|PubMed:8771792, ECO:0000303|PubMed:8824203};
DE EC=3.1.-.- {ECO:0000269|PubMed:20133613, ECO:0000269|PubMed:8771792, ECO:0000269|PubMed:8824203};
DE AltName: Full=Phosphonate monoester hydrolase {ECO:0000303|PubMed:20133613, ECO:0000303|PubMed:8771792, ECO:0000303|PubMed:8824203};
DE Short=BcPMH {ECO:0000303|PubMed:20133613};
DE EC=3.1.3.- {ECO:0000269|PubMed:20133613, ECO:0000269|PubMed:8771792, ECO:0000269|PubMed:8824203};
DE AltName: Full=Sulfuric ester hydrolase {ECO:0000305|PubMed:20133613};
DE EC=3.1.6.- {ECO:0000269|PubMed:20133613};
GN Name=pehA {ECO:0000303|PubMed:8771792, ECO:0000303|PubMed:8824203,
GN ECO:0000312|EMBL:AAC44467.1};
OS Trinickia caryophylli (Paraburkholderia caryophylli).
OC Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales;
OC Burkholderiaceae; Trinickia.
OX NCBI_TaxID=28094;
RN [1] {ECO:0000312|EMBL:AAC44467.1}
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 2-36; 134-141;
RP 161-181 AND 306-313, FUNCTION, CATALYTIC ACTIVITY, ACTIVITY REGULATION,
RP BIOPHYSICOCHEMICAL PROPERTIES, PATHWAY, SUBSTRATE SPECIFICITY, AND SUBUNIT.
RC STRAIN=PG2982 {ECO:0000303|PubMed:8824203, ECO:0000312|EMBL:AAC44467.1};
RX PubMed=8824203; DOI=10.1074/jbc.271.42.25754;
RA Dotson S.B., Smith C.E., Ling C.S., Barry G.F., Kishore G.M.;
RT "Identification, characterization, and cloning of a phosphonate monoester
RT hydrolase from Burkholderia caryophilli PG2982.";
RL J. Biol. Chem. 271:25754-25761(1996).
RN [2] {ECO:0000312|EMBL:AAC44467.1}
RP CATALYTIC ACTIVITY, AND BIOTECHNOLOGY.
RC STRAIN=PG2982 {ECO:0000303|PubMed:8771792};
RX PubMed=8771792; DOI=10.1046/j.1365-313x.1996.10020383.x;
RA Dotson S.B., Lanahan M.B., Smith A.G., Kishore G.M.;
RT "A phosphonate monoester hydrolase from Burkholderia caryophilli PG2982 is
RT useful as a conditional lethal gene in plants.";
RL Plant J. 10:383-392(1996).
RN [3] {ECO:0007744|PDB:2W8S}
RP X-RAY CRYSTALLOGRAPHY (2.40 ANGSTROMS) IN COMPLEX WITH IRON OR ZINC,
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, ACTIVITY REGULATION,
RP BIOPHYSICOCHEMICAL PROPERTIES, SUBSTRATE SPECIFICITY, REACTION MECHANISM,
RP SUBUNIT, OXOALANINE AT CYS-57, ACTIVE SITE, AND MUTAGENESIS OF CYS-57.
RC STRAIN=PG2982 {ECO:0000303|PubMed:20133613};
RX PubMed=20133613; DOI=10.1073/pnas.0903951107;
RA van Loo B., Jonas S., Babtie A.C., Benjdia A., Berteau O., Hyvonen M.,
RA Hollfelder F.;
RT "An efficient, multiply promiscuous hydrolase in the alkaline phosphatase
RT superfamily.";
RL Proc. Natl. Acad. Sci. U.S.A. 107:2740-2745(2010).
CC -!- FUNCTION: Hydrolytic enzyme with a broad substrate specificity acting
CC on phosphate diesters and phosphonate monoesters (PubMed:8824203,
CC PubMed:20133613). Hydrolyzes phosphate mono- and triesters, sulfate
CC monoesters and sulfonate monoesters (PubMed:20133613). Hydrolyzes
CC glyphosate monoesters. Does not hydrolyze DNA or cGMP (PubMed:8824203).
CC Hydrolyzes glyceryl glyphosate, but this substrate has a much lower
CC affinity than the glyphosate monoesters (PubMed:8824203,
CC PubMed:8771792). {ECO:0000269|PubMed:20133613,
CC ECO:0000269|PubMed:8771792, ECO:0000269|PubMed:8824203}.
CC -!- COFACTOR:
CC Name=Mn(2+); Xref=ChEBI:CHEBI:29035;
CC Evidence={ECO:0000305|PubMed:20133613};
CC Note=Mn(2+) is probably the active metal ion. Other metals such as
CC Zn(2+), Ca(2+) and Fe(2+) can also act as cofactors.
CC {ECO:0000303|PubMed:20133613};
CC -!- ACTIVITY REGULATION: Anions including Cl(-) and CH3COO(-), and SO4(2-)
CC salts stimulate activity 20-40% at 100 mM.
CC {ECO:0000269|PubMed:8824203}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.33 mM for phosphate monoester phenyl phosphate (at pH 7.5)
CC {ECO:0000269|PubMed:20133613};
CC KM=0.35 mM for phosphate monoester 4-nitrophenyl phosphate (at pH
CC 7.5) {ECO:0000269|PubMed:20133613};
CC KM=0.071 mM for phosphate diester diphenyl phosphate (at pH 7.5)
CC {ECO:0000269|PubMed:20133613};
CC KM=0.63 mM for phosphate diester p-nitrophenyl ethyl phosphate (at pH
CC 7.5) {ECO:0000269|PubMed:20133613};
CC KM=2.4 mM for phosphate triester paraoxon (at pH 7.5)
CC {ECO:0000269|PubMed:20133613};
CC KM=1.23 mM for phosphonate monoester phenyl phenylphosphonate (at pH
CC 7.5) {ECO:0000269|PubMed:20133613};
CC KM=0.19 mM for phosphonate monoester p-nitrophenyl phenylphosphonate
CC (at pH 7.5) {ECO:0000269|PubMed:20133613};
CC KM=58 mM for sulfate monoester phenyl sulfate (at pH 7.5)
CC {ECO:0000269|PubMed:20133613};
CC KM=68 mM for sulfate monoester p-nitrophenyl sulfate (at pH 7.5)
CC {ECO:0000269|PubMed:20133613};
CC KM=0.51 mM for sulfonate monoester phenyl phenylsulfonate (at pH 7.5)
CC {ECO:0000269|PubMed:20133613};
CC KM=0.24 mM for sulfonate monoester p-nitrophenyl phenylsulfonate (at
CC pH 7.5) {ECO:0000269|PubMed:20133613};
CC KM=49 mM for glyceryl glyphosate (at pH 9.0 and 30 degrees Celsius)
CC {ECO:0000269|PubMed:8824203};
CC KM=2.3 mM for p-nitrophenyl phenylphosphonate (at pH 9.0 and 30
CC degrees Celsius) {ECO:0000269|PubMed:8824203};
CC KM=0.9 mM for phosphate diester bis(p-nitrophenyl) phosphate (at pH
CC 9.0 and 30 degrees Celsius) {ECO:0000269|PubMed:8824203};
CC KM=4.7 mM for phosphate diester p-nitrophenyl thymidine 5'-
CC monophosphate (at pH 9.0 and 30 degrees Celsius)
CC {ECO:0000269|PubMed:8824203};
CC Vmax=8.7 umol/min/mg enzyme with p-nitrophenyl phenylphosphonate as
CC substrate {ECO:0000269|PubMed:8824203};
CC Note=kcat is 0.0021 sec(-1) with phenyl phosphate as substrate. kcat
CC is 0.077 sec(-1) with 4-nitrophenyl phosphate as substrate. kcat is
CC 2.12 sec(-1) with diphenyl phosphate as substrate. kcat is 5.8 sec(-
CC 1) with p-nitrophenyl ethyl phosphate as substrate. kcat is 0.00037
CC sec(-1) with paraoxon as substrate. kcat is 1.58 sec(-1) with phenyl
CC phenylphosphonate as substrate. kcat is 2.73 sec(-1) with p-
CC nitrophenyl phenylphosphonate as substrate. kcat is 0.001 sec(-1)
CC with phenyl sulfate as substrate. kcat is 0.4 sec(-1) with p-
CC nitrophenyl sulfate as substrate. kcat is 0.007 sec(-1) with phenyl
CC phenylsulfonate as substrate. kcat is 0.12 sec(-1) with p-nitrophenyl
CC phenylsulfonate as substrate (PubMed:20133613). kcat is 28 min(-1)
CC with glyceryl glyphosate as substrate. kcat is 400 min(-1) with p-
CC nitrophenyl phenylphosphonate as substrate. kcat is 620 min(-1) with
CC bis(p-nitrophenyl) phosphate as substrate. kcat is 89 min(-1) with p-
CC nitrophenyl thymidine 5'-monophosphate as substrate (PubMed:8824203).
CC {ECO:0000269|PubMed:20133613, ECO:0000269|PubMed:8824203};
CC pH dependence:
CC Optimum pH is 9.0. {ECO:0000269|PubMed:8824203};
CC -!- SUBUNIT: Homotetramer. {ECO:0000269|PubMed:20133613,
CC ECO:0000269|PubMed:8824203}.
CC -!- INTERACTION:
CC Q45087; Q45087: pehA; NbExp=2; IntAct=EBI-15830967, EBI-15830967;
CC -!- PTM: The conversion to 3-oxoalanine (also known as C-formylglycine,
CC FGly), of a serine or cysteine residue in prokaryotes and of a cysteine
CC residue in eukaryotes, is critical for catalytic activity (By
CC similarity). Phosphate triester hydrolytic activity is retained with
CC unmodified cysteine acting as a nucleophile (PubMed:20133613).
CC {ECO:0000255|PIRSR:PIRSR600917-51, ECO:0000269|PubMed:20133613}.
CC -!- BIOTECHNOLOGY: Has potential application in plant genetics as a
CC heterologous conditional lethal gene useful for cell ablation and
CC negative (counter) selection. {ECO:0000305|PubMed:8771792}.
CC -!- SIMILARITY: Belongs to the alkaline phosphatase superfamily.
CC {ECO:0000303|PubMed:20133613}.
CC -!- CAUTION: There is conflicting evidence for Zn(2+) acting as an active
CC metal ion. According to one report, it inhibits the phosphonate ester
CC hydrolase activity (PubMed:8824203). According to another report, it
CC acts as a cofactor (PubMed:20133613). {ECO:0000269|PubMed:20133613,
CC ECO:0000269|PubMed:8824203}.
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DR EMBL; U44852; AAC44467.1; -; Genomic_DNA.
DR PIR; T44602; T44602.
DR PDB; 2W8S; X-ray; 2.40 A; A/B/C/D=1-514.
DR PDBsum; 2W8S; -.
DR AlphaFoldDB; Q45087; -.
DR SMR; Q45087; -.
DR DIP; DIP-58542N; -.
DR EvolutionaryTrace; Q45087; -.
DR GO; GO:0042802; F:identical protein binding; IPI:IntAct.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0008484; F:sulfuric ester hydrolase activity; IEA:InterPro.
DR Gene3D; 3.40.720.10; -; 1.
DR InterPro; IPR017850; Alkaline_phosphatase_core_sf.
DR InterPro; IPR032506; DUF4976.
DR InterPro; IPR000917; Sulfatase_N.
DR Pfam; PF16347; DUF4976; 1.
DR Pfam; PF00884; Sulfatase; 1.
DR SUPFAM; SSF53649; SSF53649; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Direct protein sequencing; Hydrolase; Iron; Manganese;
KW Metal-binding; Zinc.
FT INIT_MET 1
FT /note="Removed"
FT /evidence="ECO:0000269|PubMed:8824203"
FT CHAIN 2..514
FT /note="Multifunctional alkaline phosphatase superfamily
FT protein PehA"
FT /evidence="ECO:0000305|PubMed:8824203"
FT /id="PRO_0000442718"
FT ACT_SITE 57
FT /note="Nucleophile"
FT /evidence="ECO:0000305|PubMed:20133613"
FT BINDING 12
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /evidence="ECO:0000269|PubMed:20133613,
FT ECO:0007744|PDB:2W8S"
FT BINDING 57
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /note="via 3-oxoalanine"
FT /evidence="ECO:0000269|PubMed:20133613,
FT ECO:0007744|PDB:2W8S"
FT BINDING 324
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /evidence="ECO:0000269|PubMed:20133613,
FT ECO:0007744|PDB:2W8S"
FT BINDING 325
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /evidence="ECO:0000269|PubMed:20133613,
FT ECO:0007744|PDB:2W8S"
FT MOD_RES 57
FT /note="3-oxoalanine (Cys)"
FT /evidence="ECO:0000255|PIRSR:PIRSR600917-51,
FT ECO:0000269|PubMed:20133613"
FT MUTAGEN 57
FT /note="C->A: Reduced catalytic activity against phosphate
FT mono-, di-, and triesters, phosphonate monoesters, sulfate
FT monoesters and sulfonate monoesters while KM values are
FT almost unchanged for all of them. 144-fold decrease in kcat
FT and 467-fold reduction in catalytic efficiency with
FT phosphonate monoester as substrate. 103-fold decrease in
FT kcat and 1178-fold reduction in catalytic efficiency with
FT phosphate diester as substrate."
FT /evidence="ECO:0000269|PubMed:20133613"
SQ SEQUENCE 514 AA; 58176 MW; 88503D771830A58D CRC64;
MTRKNVLLIV VDQWRADFIP HLMRAEGREP FLKTPNLDRL CREGLTFRNH VTTCVPCGPA
RASLLTGLYL MNHRAVQNTV PLDQRHLNLG KALRAIGYDP ALIGYTTTTP DPRTTSARDP
RFTVLGDIMD GFRSVGAFEP NMEGYFGWVA QNGFELPENR EDIWLPEGEH SVPGATDKPS
RIPKEFSDST FFTERALTYL KGRDGKPFFL HLGYYRPHPP FVASAPYHAM YKAEDMPAPI
RAENPDAEAA QHPLMKHYID HIRRGSFFHG AEGSGATLDE GEIRQMRATY CGLITEIDDC
LGRVFAYLDE TGQWDDTLII FTSDHGEQLG DHHLLGKIGY NAESFRIPLV IKDAGQNRHA
GQIEEGFSES IDVMPTILEW LGGETPRACD GRSLLPFLAE GKPSDWRTEL HYEFDFRDVF
YDQPQNSVQL SQDDCSLCVI EDENYKYVHF AALPPLFFDL KADPHEFSNL AGDPAYAALV
RDYAQKALSW RLSHADRTLT HYRSSPQGLT TRNH
