AMBB_PSEAE
ID AMBB_PSEAE Reviewed; 1249 AA.
AC Q9I1H0;
DT 23-FEB-2022, integrated into UniProtKB/Swiss-Prot.
DT 01-MAR-2001, sequence version 1.
DT 03-AUG-2022, entry version 134.
DE RecName: Full=AMB antimetabolite synthase AmbB {ECO:0000305};
DE EC=6.2.1.67 {ECO:0000269|PubMed:25814981};
DE AltName: Full=L-alanine--[L-alanyl-carrier protein] ligase {ECO:0000305};
DE AltName: Full=Nonribosomal peptide synthase AmbB {ECO:0000305};
GN Name=ambB {ECO:0000303|PubMed:20543073};
GN OrderedLocusNames=PA2305 {ECO:0000312|EMBL:AAG05693.1};
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 FUNCTION, AND DISRUPTION PHENOTYPE.
RC STRAIN=ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C /
RC PRS 101 / PAO1;
RX PubMed=20543073; DOI=10.1128/jb.00492-10;
RA Lee X., Fox A., Sufrin J., Henry H., Majcherczyk P., Haas D., Reimmann C.;
RT "Identification of the biosynthetic gene cluster for the Pseudomonas
RT aeruginosa antimetabolite L-2-amino-4-methoxy-trans-3-butenoic acid.";
RL J. Bacteriol. 192:4251-4255(2010).
RN [3]
RP PROPOSED FUNCTION IN IQS BIOSYNTHESIS, INDUCTION, AND DISRUPTION PHENOTYPE.
RC STRAIN=ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C /
RC PRS 101 / PAO1;
RX PubMed=23542643; DOI=10.1038/nchembio.1225;
RA Lee J., Wu J., Deng Y., Wang J., Wang C., Wang J., Chang C., Dong Y.,
RA Williams P., Zhang L.H.;
RT "A cell-cell communication signal integrates quorum sensing and stress
RT response.";
RL Nat. Chem. Biol. 9:339-343(2013).
RN [4]
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, DOMAIN, AND MUTAGENESIS OF SER-768.
RC STRAIN=ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C /
RC PRS 101 / PAO1;
RX PubMed=25814981; DOI=10.3389/fmicb.2015.00170;
RA Rojas Murcia N., Lee X., Waridel P., Maspoli A., Imker H.J., Chai T.,
RA Walsh C.T., Reimmann C.;
RT "The Pseudomonas aeruginosa antimetabolite L -2-amino-4-methoxy-trans-3-
RT butenoic acid (AMB) is made from glutamate and two alanine residues via a
RT thiotemplate-linked tripeptide precursor.";
RL Front. Microbiol. 6:170-170(2015).
RN [5]
RP CRYSTALLIZATION OF 727-1249.
RC STRAIN=ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C /
RC PRS 101 / PAO1;
RX PubMed=24598922; DOI=10.1107/s2053230x14001782;
RA Wang Y., Li D., Huan X., Zhang L., Song H.;
RT "Crystallization and preliminary X-ray crystallographic analysis of a
RT putative nonribosomal peptide synthase AmbB from Pseudomonas aeruginosa.";
RL Acta Crystallogr. F Struct. Biol. Commun. 70:339-342(2014).
CC -!- FUNCTION: Involved in the biosynthesis of the antimetabolite L-2-amino-
CC 4-methoxy-trans-3-butenoic acid (AMB), a non-proteinogenic amino acid
CC which is toxic for prokaryotes and eukaryotes (PubMed:20543073,
CC PubMed:25814981). Adenylates L-alanine and loads it onto its peptidyl
CC carrier domain via a thioester linkage to the phosphopanthetheine
CC moiety. In addition, loads activated L-Ala in trans onto the second
CC carrier domain of AmbE (PubMed:25814981). Can also activate L-Ser, Gly
CC and D-Ala, albeit to a lower extent (PubMed:25814981). The condensation
CC domain of AmbB probably condenses the activated L-Ala and the L-Glu
CC loaded on AmbE to form a L-Glu-L-Ala dipeptide at the first carrier
CC domain of AmbE (PubMed:25814981). {ECO:0000269|PubMed:20543073,
CC ECO:0000269|PubMed:25814981}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=ATP + holo-[peptidyl-carrier protein] + L-alanine = AMP +
CC diphosphate + L-alanyl-[peptidyl-carrier protein];
CC Xref=Rhea:RHEA:61800, Rhea:RHEA-COMP:11480, Rhea:RHEA-COMP:15938,
CC ChEBI:CHEBI:30616, ChEBI:CHEBI:33019, ChEBI:CHEBI:57972,
CC ChEBI:CHEBI:64479, ChEBI:CHEBI:144958, ChEBI:CHEBI:456215;
CC EC=6.2.1.67; Evidence={ECO:0000269|PubMed:25814981};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:61801;
CC Evidence={ECO:0000269|PubMed:25814981};
CC -!- COFACTOR:
CC Name=pantetheine 4'-phosphate; Xref=ChEBI:CHEBI:47942;
CC Evidence={ECO:0000269|PubMed:25814981};
CC -!- INDUCTION: Expression is regulated by the PhoR-PhoB two-component
CC system. {ECO:0000269|PubMed:23542643}.
CC -!- DOMAIN: Modular protein that contains an adenylation domain which
CC activates the alanine residue into an aminoacyl-AMP ester, a peptidyl
CC carrier protein domain which bears a phosphopantetheinyl arm to attach
CC the activated alanine and a condensation domain involved in the
CC condensation of this amino acid with a second amino acid bound at the
CC carrier protein domain of another module.
CC {ECO:0000305|PubMed:25814981}.
CC -!- DISRUPTION PHENOTYPE: Mutation abolishes AMB production
CC (PubMed:20543073, PubMed:23542643). Deletion of the gene almost
CC abolishes the production of the quorum sensing signals 2-heptyl-3-
CC hydroxy-4(1H)-quinolone (PQS) and N-butanoylhomoserine lactone (C4HSL),
CC but it has no effect on the biosynthesis of the quorum sensing signal
CC N-3-oxododecanoylhomoserine lactone (3OC12HSL) (PubMed:23542643).
CC Deletion causes a substantial reduction in the production of the
CC virulence factors pyocyanine and elastase, and it reduces bacterial
CC virulence by about 32-40% (PubMed:23542643).
CC {ECO:0000269|PubMed:20543073, ECO:0000269|PubMed:23542643}.
CC -!- SIMILARITY: Belongs to the NRP synthetase family. {ECO:0000305}.
CC -!- CAUTION: It was suggested by Lee et al that the amb cluster is involved
CC in the biosynthesis of 2-(2-hydroxyphenyl)-thiazole-4-carbaldehyde
CC (IQS), a cell-cell communication signal that modulates the production
CC of AMB through the pqs and rhl quorum sensing systems
CC (PubMed:23542643). The chemical structure of IQS indicates that this
CC compound may be assembled from salicylate and cysteine. However,
CC neither of the two peptide synthases encoded by the amb gene cluster
CC present adenylation domains with a specificity for these substrates. It
CC is thus highly implausible that IQS is specified by the amb gene
CC cluster (PubMed:25814981). {ECO:0000269|PubMed:23542643,
CC ECO:0000269|PubMed:25814981}.
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DR EMBL; AE004091; AAG05693.1; -; Genomic_DNA.
DR PIR; C83358; C83358.
DR RefSeq; NP_250995.1; NC_002516.2.
DR RefSeq; WP_003113094.1; NC_002516.2.
DR AlphaFoldDB; Q9I1H0; -.
DR SMR; Q9I1H0; -.
DR STRING; 287.DR97_6127; -.
DR PaxDb; Q9I1H0; -.
DR PRIDE; Q9I1H0; -.
DR EnsemblBacteria; AAG05693; AAG05693; PA2305.
DR GeneID; 878073; -.
DR KEGG; pae:PA2305; -.
DR PATRIC; fig|208964.12.peg.2409; -.
DR PseudoCAP; PA2305; -.
DR HOGENOM; CLU_000022_2_10_6; -.
DR InParanoid; Q9I1H0; -.
DR OMA; HHIAVDD; -.
DR PhylomeDB; Q9I1H0; -.
DR BioCyc; MetaCyc:MON-20272; -.
DR BioCyc; PAER208964:G1FZ6-2344-MON; -.
DR BRENDA; 6.2.1.67; 5087.
DR Proteomes; UP000002438; Chromosome.
DR GO; GO:0005737; C:cytoplasm; IBA:GO_Central.
DR GO; GO:0000036; F:acyl carrier activity; IBA:GO_Central.
DR GO; GO:0016874; F:ligase activity; IEA:UniProtKB-KW.
DR GO; GO:0031177; F:phosphopantetheine binding; IBA:GO_Central.
DR Gene3D; 1.10.1200.10; -; 1.
DR Gene3D; 3.30.300.30; -; 1.
DR Gene3D; 3.30.559.10; -; 1.
DR InterPro; IPR010071; AA_adenyl_domain.
DR InterPro; IPR036736; ACP-like_sf.
DR InterPro; IPR025110; AMP-bd_C.
DR InterPro; IPR045851; AMP-bd_C_sf.
DR InterPro; IPR020845; AMP-binding_CS.
DR InterPro; IPR000873; AMP-dep_Synth/Lig.
DR InterPro; IPR023213; CAT-like_dom_sf.
DR InterPro; IPR001242; Condensatn.
DR InterPro; IPR009081; PP-bd_ACP.
DR InterPro; IPR006162; Ppantetheine_attach_site.
DR Pfam; PF00501; AMP-binding; 1.
DR Pfam; PF13193; AMP-binding_C; 1.
DR Pfam; PF00668; Condensation; 1.
DR Pfam; PF00550; PP-binding; 1.
DR SUPFAM; SSF47336; SSF47336; 1.
DR TIGRFAMs; TIGR01733; AA-adenyl-dom; 1.
DR PROSITE; PS00455; AMP_BINDING; 1.
DR PROSITE; PS50075; CARRIER; 1.
DR PROSITE; PS00012; PHOSPHOPANTETHEINE; 1.
PE 1: Evidence at protein level;
KW Ligase; Phosphopantetheine; Phosphoprotein; Reference proteome.
FT CHAIN 1..1249
FT /note="AMB antimetabolite synthase AmbB"
FT /id="PRO_0000454845"
FT DOMAIN 734..809
FT /note="Carrier"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00258"
FT REGION 245..633
FT /note="Adenylation"
FT /evidence="ECO:0000305"
FT REGION 716..735
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT REGION 823..1150
FT /note="Condensation"
FT /evidence="ECO:0000305"
FT MOD_RES 768
FT /note="O-(pantetheine 4'-phosphoryl)serine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00258"
FT MUTAGEN 768
FT /note="S->A: Can activate L-Ala but cannot load it onto its
FT own carrier domain. Mutant loses the ability to make AMB."
FT /evidence="ECO:0000269|PubMed:25814981"
SQ SEQUENCE 1249 AA; 134447 MW; B189E7FD3CB2C5BA CRC64;
MQERHGLPLR SFSSGKALTA GRAVRPEVAQ ERRYLQGAPL GLELPGRIAL RDPHCAWQWF
EPEAAAEAFP AAHWLAAFLV LLGRYGNEEI TLGFPEPITV RGRQAPALLR SSYRAMESSA
ERSARLAEEL DDARRQLSAD GQERAALAGR CAVQVLAARP TASSPGWLAL VLAADGSVGL
ALRDPQYDEL RRIAGHLARL ARGLVDAQAC VGRLPWLDAD EERRLQALRS EPQAAPSRGV
LHHLFEAQAR RTPQRIAVHA ADRSLSYAEL ERESAALAVR LRAAGVAPEQ RVGVCLRRDS
GLLVGLLGVL RAGGCYVPLD PAYPEERVAY MLDDADCLLV LVDASTRERV AALGRPCLTL
EEGGDQANDL ALPASEVGAD HLAYIIYTSG STGRPKGVAI EHGSAHAFLR WAGQHYAAEE
WSGVLAATSV CFDLSVYELF GTLAEGGTLH LVENLFSLPD YPRRDEISLL NTVPSVCAAL
LALGDLPGGV RTLNLAGEPL RGHLVRQIRG QPQVRRLVNL YGPTEDTTYS TVHELDLHAE
ALDEPPIGRP LPGTTVEVLD GFEAPLPLGV AGELYLGGIG LARGYFGKPE QTAERFRVDP
GSGERRYRTG DRVRMREDGV LEHLGRLDDQ VKFNGFRIEL GEIASCLASF PGVSEACAML
TEDSAGLRRL VGYLAAPFAP PLQALNEHLG QSLPHYMLPS AFVVLAELPK TLNGKIDRKA
LPRPQATGAE PQALPSDPLE QALHQAWQAQ LGAPPRAGQG FYAAGGDSLR AVHLLATLRQ
RLSRRVPLQA FAGGPATPEA LLELLRQAAP EGDEPEPSAG AAGLSLAERR LWVAQQLAPE
DTSYNLLAHL RIVGATADAI EQALRQLLER HVALRRRVET GVDGPQPHAL AAHAVPLQRL
LASDAVHAER LLEDGVRREG ARVFDLAHEA PARLLLVVTR DSARADLLLS VHHYAFDDVS
LAVFAAELKT LLDGGRLGVL ASTPEQVAAR ERAALASGRL DRVAERWAER LLPLAKAPGA
APARPEESGG RAGQRLALPV SAAVHAACRA LAERTSVSPF SAALQAFAEV LGAELGVDDL
LVGVALAGRS RLEMQGLVGC FVNLLPLAVG LRPEQSVEWR LRQVGHDLLE LLEHQDVPLE
CVTQALRQRG ASGLPIRIAC GAHNGRAAPA VDAGVRVEAD FIPVPGARLD LTLWLEDQPQ
GWLAVWTGVS AIFDLHRIER LHQAWERRLL ANAGEPISKR MSPEGCNAS
