ODH_PSEAE
ID ODH_PSEAE Reviewed; 433 AA.
AC Q9HUX5;
DT 08-MAY-2019, integrated into UniProtKB/Swiss-Prot.
DT 01-MAR-2001, sequence version 1.
DT 03-AUG-2022, entry version 100.
DE RecName: Full=Pseudopaline synthase {ECO:0000305|PubMed:29091735};
DE EC=1.5.1.- {ECO:0000269|PubMed:29091735, ECO:0000269|PubMed:29618515};
DE AltName: Full=Opine dehydrogenase {ECO:0000303|PubMed:29091735, ECO:0000303|PubMed:29618515};
DE Short=ODH {ECO:0000303|PubMed:29091735, ECO:0000303|PubMed:29618515};
DE AltName: Full=Pseudopaline dehydrogenase {ECO:0000303|PubMed:29618515};
GN Name=cntM {ECO:0000250|UniProtKB:A0A0H2ZH12};
GN Synonyms=zrmC {ECO:0000303|PubMed:28898501};
GN OrderedLocusNames=PA4835 {ECO:0000312|EMBL:AAG08220.1};
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 [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 [2]
RP INDUCTION.
RC STRAIN=ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C /
RC PRS 101 / PAO1;
RX PubMed=26446565; DOI=10.1038/srep14644;
RA Gi M., Lee K.M., Kim S.C., Yoon J.H., Yoon S.S., Choi J.Y.;
RT "A novel siderophore system is essential for the growth of Pseudomonas
RT aeruginosa in airway mucus.";
RL Sci. Rep. 5:14644-14644(2015).
RN [3]
RP INDUCTION.
RC STRAIN=ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C /
RC PRS 101 / PAO1;
RX PubMed=28898501; DOI=10.1111/mmi.13834;
RA Mastropasqua M.C., D'Orazio M., Cerasi M., Pacello F., Gismondi A.,
RA Canini A., Canuti L., Consalvo A., Ciavardelli D., Chirullo B.,
RA Pasquali P., Battistoni A.;
RT "Growth of Pseudomonas aeruginosa in zinc poor environments is promoted by
RT a nicotianamine-related metallophore.";
RL Mol. Microbiol. 106:543-561(2017).
RN [4]
RP FUNCTION, CATALYTIC ACTIVITY, NADPH-BINDING, AND SUBSTRATE SPECIFICITY.
RC STRAIN=ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C /
RC PRS 101 / PAO1;
RX PubMed=29091735; DOI=10.1021/acs.biochem.7b00804;
RA McFarlane J.S., Lamb A.L.;
RT "Biosynthesis of an Opine Metallophore by Pseudomonas aeruginosa.";
RL Biochemistry 56:5967-5971(2017).
RN [5] {ECO:0007744|PDB:6C4N}
RP X-RAY CRYSTALLOGRAPHY (1.95 ANGSTROMS) IN COMPLEX WITH NADP, FUNCTION,
RP CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, SUBSTRATE SPECIFICITY,
RP AND SUBUNIT.
RC STRAIN=ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C /
RC PRS 101 / PAO1;
RX PubMed=29618515; DOI=10.1074/jbc.ra118.002007;
RA McFarlane J.S., Davis C.L., Lamb A.L.;
RT "Staphylopine, pseudopaline, and yersinopine dehydrogenases: A structural
RT and kinetic analysis of a new functional class of opine dehydrogenase.";
RL J. Biol. Chem. 293:8009-8019(2018).
CC -!- FUNCTION: Catalyzes the NAD(P)H-dependent reductive condensation of
CC alpha-ketoglutarate to the intermediate formed by the adjacently
CC encoded enzyme PA4836, namely (2S)-2-amino-4-{[(1S)-1-carboxy-2-(1H-
CC imidazol-4-yl)ethyl]amino}butanoate, leading to the production of
CC pseudopaline. This is the last step in the biosynthesis of the
CC metallophore pseudopaline, which is involved in the acquisition of
CC nickel and zinc, and thus enables bacterial growth inside the host,
CC where metal access is limited. Therefore, this enzyme probably
CC contributes to Pseudomonas virulence. Can use neither pyruvate nor
CC oxaloacetate in place of alpha-ketoglutarate (PubMed:29091735,
CC PubMed:29618515). Is two-fold more efficient using NADPH than NADH as
CC the electron donor (PubMed:29618515). {ECO:0000269|PubMed:29091735,
CC ECO:0000269|PubMed:29618515}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H2O + NADP(+) + pseudopaline = (2S)-2-amino-4-{[(1S)-1-
CC carboxy-2-(1H-imidazol-4-yl)ethyl]amino}butanoate + 2-oxoglutarate +
CC H(+) + NADPH; Xref=Rhea:RHEA:59784, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:16810, ChEBI:CHEBI:57783,
CC ChEBI:CHEBI:58349, ChEBI:CHEBI:143196, ChEBI:CHEBI:143198;
CC Evidence={ECO:0000269|PubMed:29091735, ECO:0000269|PubMed:29618515};
CC PhysiologicalDirection=right-to-left; Xref=Rhea:RHEA:59786;
CC Evidence={ECO:0000305|PubMed:29091735, ECO:0000305|PubMed:29618515};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H2O + NAD(+) + pseudopaline = (2S)-2-amino-4-{[(1S)-1-carboxy-
CC 2-(1H-imidazol-4-yl)ethyl]amino}butanoate + 2-oxoglutarate + H(+) +
CC NADH; Xref=Rhea:RHEA:59788, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:16810, ChEBI:CHEBI:57540, ChEBI:CHEBI:57945,
CC ChEBI:CHEBI:143196, ChEBI:CHEBI:143198;
CC Evidence={ECO:0000269|PubMed:29618515};
CC PhysiologicalDirection=right-to-left; Xref=Rhea:RHEA:59790;
CC Evidence={ECO:0000305|PubMed:29618515};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=14 uM for alpha-ketoglutarate (at pH 8.0 and 22 degrees Celsius,
CC in the presence of NADPH) {ECO:0000269|PubMed:29618515};
CC KM=61 uM for alpha-ketoglutarate (at pH 8.0 and 22 degrees Celsius,
CC in the presence of NADH) {ECO:0000269|PubMed:29618515};
CC Note=kcat is 0.42 sec(-1) with alpha-ketoglutarate as substrate,
CC using NADPH. kcat is 0.92 sec(-1) with alpha-ketoglutarate as
CC substrate, using NADH (at pH 8.0 and 22 degrees Celsius).
CC {ECO:0000269|PubMed:29618515};
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:29618515}.
CC -!- SUBCELLULAR LOCATION: Cell membrane {ECO:0000255}; Single-pass membrane
CC protein {ECO:0000255}.
CC -!- INDUCTION: Is part of the operon cntOLMI that is negatively regulated
CC by zinc level through the Zur repressor, which leads to transcriptional
CC activation of this operon under zinc depletion (PubMed:28898501).
CC Highly induced in response to airway mucus secretions (AMS) treatment
CC (PubMed:26446565). {ECO:0000269|PubMed:26446565,
CC ECO:0000269|PubMed:28898501}.
CC -!- SIMILARITY: Belongs to the staphylopine dehydrogenase family.
CC {ECO:0000305}.
CC ---------------------------------------------------------------------------
CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms
CC Distributed under the Creative Commons Attribution (CC BY 4.0) License
CC ---------------------------------------------------------------------------
DR EMBL; AE004091; AAG08220.1; -; Genomic_DNA.
DR PIR; G83042; G83042.
DR RefSeq; NP_253522.1; NC_002516.2.
DR RefSeq; WP_003112333.1; NZ_QZGE01000002.1.
DR PDB; 6C4N; X-ray; 1.95 A; A/B=1-433.
DR PDB; 6PBM; X-ray; 1.57 A; A/B=1-433.
DR PDB; 6PBN; X-ray; 1.65 A; A/B=1-433.
DR PDB; 6PBP; X-ray; 1.64 A; A/B=1-433.
DR PDB; 6PBT; X-ray; 2.18 A; A/B=1-433.
DR PDBsum; 6C4N; -.
DR PDBsum; 6PBM; -.
DR PDBsum; 6PBN; -.
DR PDBsum; 6PBP; -.
DR PDBsum; 6PBT; -.
DR AlphaFoldDB; Q9HUX5; -.
DR SMR; Q9HUX5; -.
DR STRING; 287.DR97_2185; -.
DR PaxDb; Q9HUX5; -.
DR EnsemblBacteria; AAG08220; AAG08220; PA4835.
DR GeneID; 882352; -.
DR KEGG; pae:PA4835; -.
DR PATRIC; fig|208964.12.peg.5066; -.
DR PseudoCAP; PA4835; -.
DR HOGENOM; CLU_637619_0_0_6; -.
DR OMA; RMPKEDY; -.
DR BioCyc; PAER208964:G1FZ6-4949-MON; -.
DR Proteomes; UP000002438; Chromosome.
DR GO; GO:0016021; C:integral component of membrane; IEA:UniProtKB-KW.
DR GO; GO:0005886; C:plasma membrane; IEA:UniProtKB-SubCell.
DR GO; GO:0016491; F:oxidoreductase activity; IEA:UniProtKB-KW.
DR InterPro; IPR036291; NAD(P)-bd_dom_sf.
DR InterPro; IPR016935; Opine_metallophore_DH.
DR Pfam; PF10100; Staph_opine_DH; 1.
DR PIRSF; PIRSF029692; UCP029692; 1.
DR SUPFAM; SSF51735; SSF51735; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Cell membrane; Membrane; NAD; NADP; Oxidoreductase;
KW Reference proteome; Transmembrane; Transmembrane helix.
FT CHAIN 1..433
FT /note="Pseudopaline synthase"
FT /id="PRO_0000447039"
FT TRANSMEM 7..27
FT /note="Helical"
FT /evidence="ECO:0000255"
FT ACT_SITE 219
FT /note="Proton donor/acceptor"
FT /evidence="ECO:0000250|UniProtKB:Q8CKU7"
FT BINDING 16..19
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:29618515"
FT BINDING 39..40
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:29618515"
FT BINDING 154
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:29618515"
FT BINDING 364
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:29618515"
FT STRAND 10..15
FT /evidence="ECO:0007829|PDB:6PBM"
FT HELIX 18..30
FT /evidence="ECO:0007829|PDB:6PBM"
FT STRAND 33..38
FT /evidence="ECO:0007829|PDB:6PBM"
FT HELIX 43..53
FT /evidence="ECO:0007829|PDB:6PBM"
FT STRAND 58..62
FT /evidence="ECO:0007829|PDB:6PBM"
FT HELIX 64..69
FT /evidence="ECO:0007829|PDB:6PBM"
FT STRAND 71..73
FT /evidence="ECO:0007829|PDB:6PBM"
FT STRAND 76..80
FT /evidence="ECO:0007829|PDB:6PBM"
FT HELIX 82..84
FT /evidence="ECO:0007829|PDB:6PBM"
FT STRAND 90..94
FT /evidence="ECO:0007829|PDB:6PBM"
FT HELIX 98..100
FT /evidence="ECO:0007829|PDB:6PBM"
FT HELIX 101..107
FT /evidence="ECO:0007829|PDB:6PBM"
FT HELIX 110..112
FT /evidence="ECO:0007829|PDB:6PBM"
FT STRAND 118..121
FT /evidence="ECO:0007829|PDB:6PBM"
FT HELIX 128..135
FT /evidence="ECO:0007829|PDB:6PBM"
FT STRAND 143..149
FT /evidence="ECO:0007829|PDB:6PBM"
FT STRAND 151..157
FT /evidence="ECO:0007829|PDB:6PBM"
FT STRAND 164..170
FT /evidence="ECO:0007829|PDB:6PBM"
FT STRAND 172..180
FT /evidence="ECO:0007829|PDB:6PBM"
FT HELIX 185..194
FT /evidence="ECO:0007829|PDB:6PBM"
FT TURN 195..197
FT /evidence="ECO:0007829|PDB:6PBM"
FT STRAND 199..202
FT /evidence="ECO:0007829|PDB:6PBM"
FT HELIX 206..210
FT /evidence="ECO:0007829|PDB:6PBM"
FT HELIX 214..223
FT /evidence="ECO:0007829|PDB:6PBM"
FT HELIX 226..232
FT /evidence="ECO:0007829|PDB:6PBM"
FT STRAND 235..237
FT /evidence="ECO:0007829|PDB:6PBM"
FT TURN 246..248
FT /evidence="ECO:0007829|PDB:6PBM"
FT STRAND 249..251
FT /evidence="ECO:0007829|PDB:6PBM"
FT HELIX 253..272
FT /evidence="ECO:0007829|PDB:6PBM"
FT HELIX 280..287
FT /evidence="ECO:0007829|PDB:6PBM"
FT TURN 293..295
FT /evidence="ECO:0007829|PDB:6PBM"
FT HELIX 298..302
FT /evidence="ECO:0007829|PDB:6PBM"
FT HELIX 304..306
FT /evidence="ECO:0007829|PDB:6PBM"
FT HELIX 309..322
FT /evidence="ECO:0007829|PDB:6PBM"
FT HELIX 360..377
FT /evidence="ECO:0007829|PDB:6PBM"
FT TURN 378..380
FT /evidence="ECO:0007829|PDB:6PBM"
FT HELIX 384..404
FT /evidence="ECO:0007829|PDB:6PBM"
FT HELIX 406..408
FT /evidence="ECO:0007829|PDB:6PBM"
FT HELIX 411..413
FT /evidence="ECO:0007829|PDB:6PBM"
FT HELIX 419..430
FT /evidence="ECO:0007829|PDB:6PBM"
SQ SEQUENCE 433 AA; 47715 MW; DB6BCAAC0BFE00ED CRC64;
MNAADESLGN VLLVGLGAVA IQVALDLRRH GAGRLGALNH PGRRSQRIAE ALARGACLQL
EGQGQHRWLS GNAALDVFHQ DPAELRDDWQ TLVLCVPADS YLDVVRGLPW ERLGGVRTLL
LVSAFIGANL LVRSALPAGC QATVLSLSSY YAATKVIDET QPLRALTKAV KRRVYLGSSR
PDCPARETWR RVLAGSGVEV VPLATPEAAE GRNVTTYVHS PFFLGEFALA RILSEQGPPG
FMYKLYPEGP ITPGAIGAMR RLWCELSELL RRMGAEPLNL LRFLNDDNYP VHETMLPRAS
IDGFAEAGAE RQEYLLFVRY AALLVDPFSP ADEQGRHFDF SAVPFRRVSR DEDGLWRLPR
VPLEDYRKLA LIVALAAHFD LAMPQARSLL ASYENAVSRF IDCQGASQCH PSLYPIDSRP
AADAIYRQWC STC
