ID   LPLA_ECOLI              Reviewed;         338 AA.
AC   P32099; Q2M5T1;
DT   01-OCT-1993, integrated into UniProtKB/Swiss-Prot.
DT   23-JAN-2007, sequence version 3.
DT   02-SEP-2026, entry version 193.
DE   RecName: Full=Lipoate-protein ligase A;
DE            EC=6.3.1.20;
DE   AltName: Full=Lipoate--protein ligase;
GN   Name=lplA; Synonyms=yjjF; OrderedLocusNames=b4386, JW4349;
OS   Escherichia coli (strain K12).
OC   Bacteria; Pseudomonadati; Pseudomonadota; Gammaproteobacteria;
OC   Enterobacterales; Enterobacteriaceae; Escherichia.
OX   NCBI_TaxID=83333;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 2-26, AND
RP   CHARACTERIZATION.
RC   STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
RX   PubMed=8206909; DOI=10.1016/s0021-9258(17)33977-7;
RA   Morris T.W., Reed K.E., Cronan J.E. Jr.;
RT   "Identification of the gene encoding lipoate-protein ligase A of
RT   Escherichia coli. Molecular cloning and characterization of the lplA gene
RT   and gene product.";
RL   J. Biol. Chem. 269:16091-16100(1994).
RN   [2]
RP   NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC   STRAIN=K12 / MG1655 / ATCC 47076;
RX   PubMed=7610040; DOI=10.1093/nar/23.12.2105;
RA   Burland V.D., Plunkett G. III, Sofia H.J., Daniels D.L., Blattner F.R.;
RT   "Analysis of the Escherichia coli genome VI: DNA sequence of the region
RT   from 92.8 through 100 minutes.";
RL   Nucleic Acids Res. 23:2105-2119(1995).
RN   [3]
RP   NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC   STRAIN=K12 / MG1655 / ATCC 47076;
RX   PubMed=9278503; DOI=10.1126/science.277.5331.1453;
RA   Blattner F.R., Plunkett G. III, Bloch C.A., Perna N.T., Burland V.,
RA   Riley M., Collado-Vides J., Glasner J.D., Rode C.K., Mayhew G.F.,
RA   Gregor J., Davis N.W., Kirkpatrick H.A., Goeden M.A., Rose D.J., Mau B.,
RA   Shao Y.;
RT   "The complete genome sequence of Escherichia coli K-12.";
RL   Science 277:1453-1462(1997).
RN   [4]
RP   NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC   STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
RX   PubMed=16738553; DOI=10.1038/msb4100049;
RA   Hayashi K., Morooka N., Yamamoto Y., Fujita K., Isono K., Choi S.,
RA   Ohtsubo E., Baba T., Wanner B.L., Mori H., Horiuchi T.;
RT   "Highly accurate genome sequences of Escherichia coli K-12 strains MG1655
RT   and W3110.";
RL   Mol. Syst. Biol. 2:E1-E5(2006).
RN   [5]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 1-68.
RC   STRAIN=K12;
RX   PubMed=2684780; DOI=10.1016/0378-1119(89)90047-4;
RA   Neuwald A.F., Stauffer G.V.;
RT   "An Escherichia coli membrane protein with a unique signal sequence.";
RL   Gene 82:219-228(1989).
RN   [6]
RP   PARTIAL PROTEIN SEQUENCE, FUNCTION, SUBSTRATE SPECIFICITY, SUBUNIT, AND
RP   BIOPHYSICOCHEMICAL PROPERTIES.
RC   STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
RX   PubMed=7639702; DOI=10.1042/bj3090853;
RA   Green D.E., Morris T.W., Green J., Cronan J.E. Jr., Guest J.R.;
RT   "Purification and properties of the lipoate protein ligase of Escherichia
RT   coli.";
RL   Biochem. J. 309:853-862(1995).
RN   [7]
RP   VARIANT SER-74.
RX   PubMed=8002607; DOI=10.1128/jb.177.1.1-10.1995;
RA   Morris T.W., Reed K.E., Cronan J.E. Jr.;
RT   "Lipoic acid metabolism in Escherichia coli: the lplA and lipB genes define
RT   redundant pathways for ligation of lipoyl groups to apoprotein.";
RL   J. Bacteriol. 177:1-10(1995).
RN   [8]
RP   CHARACTERIZATION.
RC   STRAIN=K12 / JK1;
RX   PubMed=12591875; DOI=10.1128/jb.185.5.1582-1589.2003;
RA   Jordan S.W., Cronan J.E. Jr.;
RT   "The Escherichia coli lipB gene encodes lipoyl (octanoyl)-acyl carrier
RT   protein:protein transferase.";
RL   J. Bacteriol. 185:1582-1589(2003).
RN   [9]
RP   X-RAY CRYSTALLOGRAPHY (2.4 ANGSTROMS), AND MUTAGENESIS OF SER-73 AND
RP   ARG-141.
RX   PubMed=16043486; DOI=10.1074/jbc.m505010200;
RA   Fujiwara K., Toma S., Okamura-Ikeda K., Motokawa Y., Nakagawa A.,
RA   Taniguchi H.;
RT   "Crystal structure of lipoate-protein ligase A from Escherichia coli.
RT   Determination of the lipoic acid-binding site.";
RL   J. Biol. Chem. 280:33645-33651(2005).
CC   -!- FUNCTION: Catalyzes both the ATP-dependent activation of exogenously
CC       supplied lipoate to lipoyl-AMP and the transfer of the activated lipoyl
CC       onto the lipoyl domains of lipoate-dependent enzymes. Is also able to
CC       catalyze very poorly the transfer of lipoyl and octanoyl moiety from
CC       their acyl carrier protein. {ECO:0000269|PubMed:7639702}.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=L-lysyl-[lipoyl-carrier protein] + (R)-lipoate + ATP = N(6)-
CC         [(R)-lipoyl]-L-lysyl-[lipoyl-carrier protein] + AMP + diphosphate +
CC         H(+); Xref=Rhea:RHEA:49288, Rhea:RHEA-COMP:10500, Rhea:RHEA-
CC         COMP:10502, ChEBI:CHEBI:15378, ChEBI:CHEBI:29969, ChEBI:CHEBI:30616,
CC         ChEBI:CHEBI:33019, ChEBI:CHEBI:83088, ChEBI:CHEBI:83099,
CC         ChEBI:CHEBI:456215; EC=6.3.1.20;
CC   -!- ACTIVITY REGULATION: 6-seleno-octanoate, 8-thio-octanoate and 8-seleno-
CC       octanoate caused 100%, 50% and 63% inhibition respectively. Inhibited
CC       by Cu(2+).
CC   -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC       Kinetic parameters:
CC         KM=1.9 uM for ATP {ECO:0000269|PubMed:7639702};
CC         KM=1.7 uM for D,L-lipoic acid {ECO:0000269|PubMed:7639702};
CC         KM=152 uM for magnesium ion {ECO:0000269|PubMed:7639702};
CC         Vmax=40 nmol/min/mg enzyme toward ATP {ECO:0000269|PubMed:7639702};
CC         Vmax=24 nmol/min/mg enzyme toward D,L-lipoic acid
CC         {ECO:0000269|PubMed:7639702};
CC       pH dependence:
CC         Most active from pH 5.5 to 8.0. Inactive below pH 4.3.
CC         {ECO:0000269|PubMed:7639702};
CC   -!- PATHWAY: Protein modification; protein lipoylation via exogenous
CC       pathway; protein N(6)-(lipoyl)lysine from lipoate: step 1/2.
CC   -!- PATHWAY: Protein modification; protein lipoylation via exogenous
CC       pathway; protein N(6)-(lipoyl)lysine from lipoate: step 2/2.
CC   -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:7639702}.
CC   -!- SUBCELLULAR LOCATION: Cytoplasm.
CC   -!- MISCELLANEOUS: In the transfer reaction, the free carboxyl group of
CC       lipoic acid is attached via an amide linkage to the epsilon-amino group
CC       of a specific lysine residue of lipoyl domains of lipoate-dependent
CC       enzymes.
CC   -!- SIMILARITY: Belongs to the LplA family. {ECO:0000305}.
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DR   EMBL; L27665; AAA21740.1; -; Genomic_DNA.
DR   EMBL; U14003; AAA97282.1; -; Genomic_DNA.
DR   EMBL; U00096; AAC77339.1; -; Genomic_DNA.
DR   EMBL; AP009048; BAE78375.1; -; Genomic_DNA.
DR   EMBL; X03046; CAA26854.1; -; Genomic_DNA.
DR   PIR; A54035; A54035.
DR   RefSeq; NP_418803.1; NC_000913.3.
DR   RefSeq; WP_000105884.1; NZ_CP009789.1.
DR   PDB; 1X2G; X-ray; 2.40 A; A/B/C=2-338.
DR   PDB; 1X2H; X-ray; 2.91 A; A/B/C=2-338.
DR   PDB; 3A7A; X-ray; 3.10 A; A/C=2-338.
DR   PDB; 3A7R; X-ray; 2.05 A; A=2-338.
DR   PDB; 4TVW; X-ray; 3.50 A; A/B/C/D=1-338.
DR   PDB; 4TVY; X-ray; 2.15 A; A/B=1-338.
DR   AlphaFoldDB; P32099; -.
DR   SMR; P32099; -.
DR   BioGRID; 4262787; 26.
DR   DIP; DIP-10119N; -.
DR   FunCoup; P32099; 620.
DR   IntAct; P32099; 3.
DR   STRING; 511145.b4386; -.
DR   jPOST; P32099; -.
DR   PaxDb; 511145-b4386; -.
DR   DNASU; 944865; -.
DR   EnsemblBacteria; AAC77339; AAC77339; b4386.
DR   GeneID; 944865; -.
DR   KEGG; ecj:JW4349; -.
DR   KEGG; eco:b4386; -.
DR   KEGG; ecoc:C3026_23700; -.
DR   PATRIC; fig|1411691.4.peg.2299; -.
DR   EchoBASE; EB1744; -.
DR   eggNOG; COG0095; Bacteria.
DR   HOGENOM; CLU_022986_0_1_6; -.
DR   InParanoid; P32099; -.
DR   OMA; RYQNWDW; -.
DR   OrthoDB; 9787898at2; -.
DR   PhylomeDB; P32099; -.
DR   BioCyc; EcoCyc:EG11796-MONOMER; -.
DR   BioCyc; MetaCyc:EG11796-MONOMER; -.
DR   BRENDA; 6.3.1.20; 2026.
DR   UniPathway; UPA00537; UER00594.
DR   UniPathway; UPA00537; UER00595.
DR   EvolutionaryTrace; P32099; -.
DR   PRO; PR:P32099; -.
DR   Proteomes; UP000000625; Chromosome.
DR   GO; GO:0005737; C:cytoplasm; IBA:GO_Central.
DR   GO; GO:0005829; C:cytosol; IDA:EcoCyc.
DR   GO; GO:0016979; F:lipoate-protein ligase activity; IDA:CAFA.
DR   GO; GO:0017118; F:lipoyltransferase activity; IDA:CAFA.
DR   GO; GO:0009249; P:protein lipoylation; IDA:CAFA.
DR   CDD; cd16435; BPL_LplA_LipB; 1.
DR   FunFam; 3.30.390.50:FF:000002; Lipoate-protein ligase A; 1.
DR   FunFam; 3.30.930.10:FF:000024; Lipoate-protein ligase A; 1.
DR   Gene3D; 3.30.930.10; Bira Bifunctional Protein, Domain 2; 1.
DR   Gene3D; 3.30.390.50; CO dehydrogenase flavoprotein, C-terminal domain; 1.
DR   HAMAP; MF_01602; LplA; 1.
DR   InterPro; IPR045864; aa-tRNA-synth_II/BPL/LPL.
DR   InterPro; IPR004143; BPL_LPL_catalytic.
DR   InterPro; IPR023741; Lipoate_ligase_A.
DR   InterPro; IPR019491; Lipoate_protein_ligase_C.
DR   InterPro; IPR004562; LipoylTrfase_LipoateP_Ligase.
DR   NCBIfam; TIGR00545; lipoyltrans; 1.
DR   PANTHER; PTHR12561; LIPOATE-PROTEIN LIGASE; 1.
DR   PANTHER; PTHR12561:SF3; LIPOYLTRANSFERASE 1, MITOCHONDRIAL; 1.
DR   Pfam; PF10437; Lip_prot_lig_C; 1.
DR   Pfam; PF21948; LplA-B_cat; 1.
DR   SUPFAM; SSF55681; Class II aaRS and biotin synthetases; 1.
DR   SUPFAM; SSF82649; SufE/NifU; 1.
DR   PROSITE; PS51733; BPL_LPL_CATALYTIC; 1.
DR   PDBsum; 1X2G; -.
DR   PDBsum; 1X2H; -.
DR   PDBsum; 3A7A; -.
DR   PDBsum; 3A7R; -.
DR   PDBsum; 4TVW; -.
DR   PDBsum; 4TVY; -.
PE   1: Evidence at protein level;
KW   3D-structure; ATP-binding; Cytoplasm; Direct protein sequencing; Ligase;
KW   Nucleotide-binding; Reference proteome.
FT   INIT_MET        1
FT                   /note="Removed"
FT                   /evidence="ECO:0000269|PubMed:8206909"
FT   CHAIN           2..338
FT                   /note="Lipoate-protein ligase A"
FT                   /id="PRO_0000209564"
FT   DOMAIN          29..216
FT                   /note="BPL/LPL catalytic"
FT                   /evidence="ECO:0000255|PROSITE-ProRule:PRU01067"
FT   BINDING         71
FT                   /ligand="ATP"
FT                   /ligand_id="ChEBI:CHEBI:30616"
FT                   /evidence="ECO:0000250"
FT   BINDING         76..79
FT                   /ligand="ATP"
FT                   /ligand_id="ChEBI:CHEBI:30616"
FT                   /evidence="ECO:0000250"
FT   BINDING         134
FT                   /ligand="(R)-lipoate"
FT                   /ligand_id="ChEBI:CHEBI:83088"
FT                   /evidence="ECO:0000250"
FT   BINDING         134
FT                   /ligand="ATP"
FT                   /ligand_id="ChEBI:CHEBI:30616"
FT                   /evidence="ECO:0000250"
FT   VARIANT         74
FT                   /note="G -> S (in lplA1 or slr1; selenolipoate resistance
FT                   mutation)"
FT                   /evidence="ECO:0000269|PubMed:8002607"
FT   MUTAGEN         73
FT                   /note="S->A: 20-fold decrease in affinity for ATP."
FT                   /evidence="ECO:0000269|PubMed:16043486"
FT   MUTAGEN         141
FT                   /note="R->A: More than 10-fold reduction in Vmax."
FT                   /evidence="ECO:0000269|PubMed:16043486"
FT   STRAND          4..9
FT                   /evidence="ECO:0007829|PDB:3A7R"
FT   HELIX           14..27
FT                   /evidence="ECO:0007829|PDB:3A7R"
FT   STRAND          33..38
FT                   /evidence="ECO:0007829|PDB:3A7R"
FT   STRAND          41..46
FT                   /evidence="ECO:0007829|PDB:3A7R"
FT   HELIX           52..55
FT                   /evidence="ECO:0007829|PDB:3A7R"
FT   HELIX           58..63
FT                   /evidence="ECO:0007829|PDB:3A7R"
FT   STRAND          67..70
FT                   /evidence="ECO:0007829|PDB:3A7R"
FT   STRAND          78..80
FT                   /evidence="ECO:0007829|PDB:3A7R"
FT   STRAND          84..92
FT                   /evidence="ECO:0007829|PDB:3A7R"
FT   TURN            93..95
FT                   /evidence="ECO:0007829|PDB:3A7R"
FT   HELIX           98..111
FT                   /evidence="ECO:0007829|PDB:3A7R"
FT   STRAND          116..119
FT                   /evidence="ECO:0007829|PDB:3A7R"
FT   TURN            120..122
FT                   /evidence="ECO:0007829|PDB:3A7R"
FT   STRAND          123..126
FT                   /evidence="ECO:0007829|PDB:3A7R"
FT   STRAND          133..142
FT                   /evidence="ECO:0007829|PDB:3A7R"
FT   STRAND          144..156
FT                   /evidence="ECO:0007829|PDB:3A7R"
FT   HELIX           159..165
FT                   /evidence="ECO:0007829|PDB:3A7R"
FT   HELIX           168..170
FT                   /evidence="ECO:0007829|PDB:3A7R"
FT   STRAND          172..175
FT                   /evidence="ECO:0007829|PDB:3A7R"
FT   HELIX           178..180
FT                   /evidence="ECO:0007829|PDB:3A7R"
FT   HELIX           188..190
FT                   /evidence="ECO:0007829|PDB:3A7R"
FT   HELIX           197..212
FT                   /evidence="ECO:0007829|PDB:3A7R"
FT   STRAND          219..221
FT                   /evidence="ECO:0007829|PDB:3A7R"
FT   STRAND          223..225
FT                   /evidence="ECO:0007829|PDB:3A7A"
FT   HELIX           232..240
FT                   /evidence="ECO:0007829|PDB:3A7R"
FT   HELIX           242..245
FT                   /evidence="ECO:0007829|PDB:3A7R"
FT   TURN            246..248
FT                   /evidence="ECO:0007829|PDB:3A7R"
FT   STRAND          252..260
FT                   /evidence="ECO:0007829|PDB:3A7R"
FT   STRAND          263..272
FT                   /evidence="ECO:0007829|PDB:3A7R"
FT   STRAND          275..283
FT                   /evidence="ECO:0007829|PDB:3A7R"
FT   HELIX           289..297
FT                   /evidence="ECO:0007829|PDB:3A7R"
FT   TURN            298..300
FT                   /evidence="ECO:0007829|PDB:3A7R"
FT   STRAND          302..304
FT                   /evidence="ECO:0007829|PDB:3A7R"
FT   HELIX           305..314
FT                   /evidence="ECO:0007829|PDB:3A7R"
FT   TURN            315..318
FT                   /evidence="ECO:0007829|PDB:3A7R"
FT   HELIX           320..322
FT                   /evidence="ECO:0007829|PDB:3A7R"
FT   HELIX           323..337
FT                   /evidence="ECO:0007829|PDB:3A7R"
SQ   SEQUENCE   338 AA;  37926 MW;  102788082E182A6F CRC64;
     MSTLRLLISD SYDPWFNLAV EECIFRQMPA TQRVLFLWRN ADTVVIGRAQ NPWKECNTRR
     MEEDNVRLAR RSSGGGAVFH DLGNTCFTFM AGKPEYDKTI STSIVLNALN ALGVSAEASG
     RNDLVVKTVE GDRKVSGSAY RETKDRGFHH GTLLLNADLS RLANYLNPDK KKLAAKGITS
     VRSRVTNLTE LLPGITHEQV CEAITEAFFA HYGERVEAEI ISPNKTPDLP NFAETFARQS
     SWEWNFGQAP AFSHLLDERF TWGGVELHFD VEKGHITRAQ VFTDSLNPAP LEALAGRLQG
     CLYRADMLQQ ECEALLVDFP EQEKELRELS AWMAGAVR
//