AMBE_PSEAE
ID AMBE_PSEAE Reviewed; 2124 AA.
AC Q9I1H3;
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 AmbE {ECO:0000305};
DE EC=6.2.1.68 {ECO:0000269|PubMed:25814981};
DE AltName: Full=L-glutamate--[L-glutamyl-carrier protein] ligase {ECO:0000305};
DE AltName: Full=Nonribosomal peptide synthase AmbE {ECO:0000305};
GN Name=ambE {ECO:0000303|PubMed:20543073};
GN OrderedLocusNames=PA2302 {ECO:0000312|EMBL:AAG05690.1};
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 ASP-644;
RP LYS-1230; SER-1286 AND SER-1819.
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).
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-glutamate and loads it onto its first
CC peptidyl carrier domain via a thioester linkage to the
CC phosphopanthetheine moiety (PubMed:25814981). The second peptidyl
CC carrier domain is loaded with a L-alanine activated by AmbB
CC (PubMed:25814981). After formation by AmbB of the L-Glu-L-Ala dipeptide
CC at the first carrier domain of AmbE, the condensation domain of AmbE
CC probably condenses this dipeptide with the L-Ala residue attached at
CC the second carrier domain of AmbE to give the L-Ala-L-Glu-L-Ala
CC tripeptide. The central amino acid, L-Glu, would then undergo a series
CC of modifications to be converted into AMB while the two flanking L-Ala
CC residues remain in place (PubMed:25814981). Finally, the L-Ala-AMB-L-
CC Ala tripeptide is probably released by thioester cleavage via the
CC thioester domain of AmbE (PubMed:25814981).
CC {ECO:0000269|PubMed:20543073, ECO:0000269|PubMed:25814981}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=ATP + holo-[peptidyl-carrier protein] + L-glutamate = AMP +
CC diphosphate + L-glutamyl-[peptidyl-carrier protein];
CC Xref=Rhea:RHEA:62452, Rhea:RHEA-COMP:11480, Rhea:RHEA-COMP:16100,
CC ChEBI:CHEBI:29985, ChEBI:CHEBI:30616, ChEBI:CHEBI:33019,
CC ChEBI:CHEBI:64479, ChEBI:CHEBI:144960, ChEBI:CHEBI:456215;
CC EC=6.2.1.68; Evidence={ECO:0000269|PubMed:25814981};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:62453;
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 glutamate residue into an aminoacyl-AMP ester, a
CC methyltransferase domain, a first peptidyl carrier protein domain which
CC bears a phosphopantetheinyl arm to attach the activated L-glutamate, a
CC condensation domain involved in the condensation of this amino acid
CC with a second amino acid bound at the carrier protein domain of another
CC module, a second peptidyl carrier protein domain which bears a
CC phosphopantetheinyl arm to attach a L-alanine activated by AmbB and a
CC thioesterase domain that may release the newly synthesized peptide from
CC the enzyme. {ECO:0000305|PubMed:25814981}.
CC -!- DISRUPTION PHENOTYPE: Mutation abolishes AMB production
CC (PubMed:20543073). Deletion of the gene causes a substantial reduction
CC in the production of the quorum sensing signal 2-heptyl-3-hydroxy-
CC 4(1H)-quinolone (PQS) (PubMed:23542643). {ECO:0000269|PubMed:20543073,
CC 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; AAG05690.1; -; Genomic_DNA.
DR PIR; H83357; H83357.
DR RefSeq; NP_250992.1; NC_002516.2.
DR RefSeq; WP_010895606.1; NZ_QZGE01000036.1.
DR AlphaFoldDB; Q9I1H3; -.
DR SMR; Q9I1H3; -.
DR STRING; 287.DR97_6131; -.
DR ESTHER; pseae-PA2302; Thioesterase.
DR PaxDb; Q9I1H3; -.
DR PRIDE; Q9I1H3; -.
DR EnsemblBacteria; AAG05690; AAG05690; PA2302.
DR GeneID; 878274; -.
DR KEGG; pae:PA2302; -.
DR PATRIC; fig|208964.12.peg.2406; -.
DR PseudoCAP; PA2302; -.
DR HOGENOM; CLU_236504_0_0_6; -.
DR InParanoid; Q9I1H3; -.
DR OMA; FSEWANE; -.
DR PhylomeDB; Q9I1H3; -.
DR BioCyc; MetaCyc:MON-20273; -.
DR BioCyc; PAER208964:G1FZ6-2341-MON; -.
DR BRENDA; 6.2.1.68; 5087.
DR Proteomes; UP000002438; Chromosome.
DR GO; GO:0016874; F:ligase activity; IEA:UniProtKB-KW.
DR GO; GO:0031177; F:phosphopantetheine binding; IEA:InterPro.
DR GO; GO:0043604; P:amide biosynthetic process; IEA:UniProt.
DR GO; GO:1901566; P:organonitrogen compound biosynthetic process; IEA:UniProt.
DR GO; GO:0044550; P:secondary metabolite biosynthetic process; IEA:UniProt.
DR Gene3D; 1.10.1200.10; -; 2.
DR Gene3D; 3.30.300.30; -; 2.
DR Gene3D; 3.30.559.10; -; 2.
DR Gene3D; 3.40.50.12780; -; 1.
DR Gene3D; 3.40.50.150; -; 1.
DR Gene3D; 3.40.50.1820; -; 1.
DR InterPro; IPR010071; AA_adenyl_domain.
DR InterPro; IPR029058; AB_hydrolase.
DR InterPro; IPR036736; ACP-like_sf.
DR InterPro; IPR045851; AMP-bd_C_sf.
DR InterPro; IPR020845; AMP-binding_CS.
DR InterPro; IPR000873; AMP-dep_Synth/Lig.
DR InterPro; IPR042099; ANL_N_sf.
DR InterPro; IPR023213; CAT-like_dom_sf.
DR InterPro; IPR001242; Condensatn.
DR InterPro; IPR006342; FkbM_mtfrase.
DR InterPro; IPR020806; PKS_PP-bd.
DR InterPro; IPR020802; PKS_thioesterase.
DR InterPro; IPR009081; PP-bd_ACP.
DR InterPro; IPR006162; Ppantetheine_attach_site.
DR InterPro; IPR029063; SAM-dependent_MTases_sf.
DR InterPro; IPR001031; Thioesterase.
DR Pfam; PF00501; AMP-binding; 1.
DR Pfam; PF00668; Condensation; 1.
DR Pfam; PF05050; Methyltransf_21; 1.
DR Pfam; PF00550; PP-binding; 2.
DR Pfam; PF00975; Thioesterase; 1.
DR SMART; SM00823; PKS_PP; 2.
DR SMART; SM00824; PKS_TE; 1.
DR SUPFAM; SSF47336; SSF47336; 2.
DR SUPFAM; SSF53335; SSF53335; 1.
DR SUPFAM; SSF53474; SSF53474; 1.
DR TIGRFAMs; TIGR01733; AA-adenyl-dom; 1.
DR TIGRFAMs; TIGR01444; fkbM_fam; 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; Repeat.
FT CHAIN 1..2124
FT /note="AMB antimetabolite synthase AmbE"
FT /id="PRO_0000454846"
FT DOMAIN 1251..1325
FT /note="Carrier 1"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00258"
FT DOMAIN 1785..1859
FT /note="Carrier 2"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00258"
FT REGION 456..847
FT /note="Adenylation"
FT /evidence="ECO:0000305"
FT REGION 950..1147
FT /note="Methyltransferase"
FT /evidence="ECO:0000305"
FT REGION 1359..1780
FT /note="Condensation"
FT /evidence="ECO:0000305"
FT REGION 1886..2107
FT /note="Thioesterase"
FT /evidence="ECO:0000305"
FT MOD_RES 1286
FT /note="O-(pantetheine 4'-phosphoryl)serine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00258"
FT MOD_RES 1819
FT /note="O-(pantetheine 4'-phosphoryl)serine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00258"
FT MUTAGEN 644
FT /note="D->A: Cannot load L-glutamate; when associated with
FT T-1230."
FT /evidence="ECO:0000269|PubMed:25814981"
FT MUTAGEN 1230
FT /note="K->T: Cannot load L-glutamate; when associated with
FT A-644."
FT /evidence="ECO:0000269|PubMed:25814981"
FT MUTAGEN 1286
FT /note="S->A: Cannot load L-glutamate. Mutant loses the
FT ability to make AMB."
FT /evidence="ECO:0000269|PubMed:25814981"
FT MUTAGEN 1819
FT /note="S->A: Can still load L-glutamate. Mutant loses the
FT ability to make AMB."
FT /evidence="ECO:0000269|PubMed:25814981"
SQ SEQUENCE 2124 AA; 229044 MW; EE5018845F69E93E CRC64;
MSASEDLQSA VQPAASEALE GFPLSPLQTR AWRRHAERPE NTVVGVRLHA PADPVATLER
LRRALDGEAQ LRVAYRTMPG MSLPVQVLDG RAADLLVERL PGDGDWAGRF ARESARLAAS
PLGGEGQPVL ALGLLLDAAG ETLQGLLLAA PAFVVDAASL VALLRRGLGP AGQASADEGD
EALLFQHFSE WANEALAGED GESASGYWRE QAAVAAESPL ALADDLGEGE WTARRLLPRA
LLERLAANGL PEAAALLAWT QVAGQFQGDE GLPLEMARLV SGRLFNEFAE LAGPFAGVAP
LCLENVRAGS VGERLDALQA AILAQEEAAA LRDPFAPDWP LAELGFAWLA GELDGAGVAE
LDCRQPPLGG FLELQVLPHG EGRLASLRVR RDHDGTLAGR LLDAWVECLE SIAADRQLPL
AGLPLIGAAE RERYQAWQGE RVEPAPVESL VAAFDLRAAL QPQAPALLDA HGSLDFATLR
ARSEAVAEAL LAAGVRPGQA VAVMTGRNRE AIVALLGVMR AAAVYTPVNP EFPAARVERM
REAGGIVFAL ADAECAGRAR EAFAGACLDL STLPLAGSGM SLPAPGGRDA AYMIFTSGTS
GQPKGVVVEH ASALNLSQAL ARTVYANVVG EGLRVTVNAP FSFDSSIKQI LQLLSGHCLV
LVPQEVRSDP QRMLGFLEER RIDVLDCTPS LFRLLLQAGL DDAHPALPGR ILVGGERFDE
ASWEVAAGWR RCQVFNLYGP TEATVNASLA RVAEHARPTI GRALANVDLH VVDGLGRRKT
RGASGELWIG GAGVARGYAG DAGEAAGRFV EEGWPGSGRL YRSGDLVRWR ADGCLEFLGR
IDEQVKINGY RIELGEIRSA LLEHPAVGEA AVLTDEADAA EPGADRRIVA FVTAAEETAD
ESWLEVDLPS GHRVAGLNLN ETEYVYQEIF VDEVYSRDGI VLPPDAVVLD VGANIGLFSL
YIASRAPRAR VVAFEPLAPI RRRLEANLGR YAPQVEVFGI GLSDAEREET FTYYPGYSTF
SGIAEYADAS GERDVIRRYL SNQGEEGGAN LLLDNIDEIL DDRLRAEAHR CRLRRLDQVI
GELGLERIDL LKIDVQRAEM DVLLGLDDAA LAKVRQIVLE VHDKRDGATA GRADALSDLL
RRHGFEVSIR QDALLEGTDR YNCYAVRPGY AESLAERIDW RALAPRPAAA LGGELSEQAL
RGFLEARLPA YMLPSRIARV ERLPLTAEGK LDRRALLAAL AAEAAAQTLE APANATEAAL
LEIWKSVLKR PAIGVSDNFF QVGGDSIRLI QMQVMAREAG LAFTLRDVFN HQSIRELARL
LAAPASPADA LGTSAPQSLE PFALLSAAER KRLPEGLDDA YPMTSLQQGM LLQSEASGDP
RLLHNVVLHE VHGRLDGELL ARAWAILIGR HAILRTGFDL HGGQVPLQWV HPATAVAAEV
PVHDLCGLDG ETRRLRLRAW IEEEQATPFD WSRPPLVRLA ALALDERRFA LGVAEHHSVL
DGWSLQSLVD ELLAVYADLL AGVVAREAEA PAVGFRDYVA LEREAEANAA SALFWLDYLA
GARYRPLPGL AEEGPRRMAA VRVDVPADSL SRLRALAERS GLPLRSLLLA AHGRALCRFS
DADEVVTGFV SHGRPEEPGA DRLLGLFLNT LPCRLSASVD LLDSARRAFD YERASLEHRR
HPLAAIRRRN RELRLDSLFN FVDFHQDDAA PAGVRHGGIL DQVVVDVDVP LAVDFEVAGE
RLEVGFQYAA GRFPAERAEA LAGAYREALL ALLGDPVQPP AAAQAEDSVE LRRVLKVLSR
VLGRPLAADQ GFASAGGHSL LGVQAIAELR RLTGRQLSLG LLQGDPDARE VVRRCHAADA
PPLPPATERA RALWLQRSGS AQPRLRLIAL PPAGGNAGTF RGWDARLPAD VELLAIQYPG
RQERQDEPFV TDVEAMLCAI DDALLPLLDR PFALIGASLG GMLAYELAAR LESLHGLRAR
QLFVISSRAP GPDLEYPRFH AMGDAELLRT LREYDVLPLE VLDDPELREI SLATLRADSR
LAADYRYRPR EPLAIPITAI LGEQDPGVSR VAIDGWRRHA SRYELETLAG GHGLVVTAAE
EVCAILRQRL APDVPGGVPA NLAT
