LPLA_ECOLI
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 03-AUG-2022, entry version 177.
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; Proteobacteria; Gammaproteobacteria; Enterobacterales;
OC 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=(R)-lipoate + ATP + L-lysyl-[lipoyl-carrier protein] = (R)-
CC N(6)-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_LN832404.1.
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 PDBsum; 1X2G; -.
DR PDBsum; 1X2H; -.
DR PDBsum; 3A7A; -.
DR PDBsum; 3A7R; -.
DR PDBsum; 4TVW; -.
DR PDBsum; 4TVY; -.
DR AlphaFoldDB; P32099; -.
DR SMR; P32099; -.
DR BioGRID; 4262787; 24.
DR DIP; DIP-10119N; -.
DR IntAct; P32099; 3.
DR STRING; 511145.b4386; -.
DR jPOST; P32099; -.
DR PaxDb; P32099; -.
DR PRIDE; P32099; -.
DR DNASU; 944865; -.
DR EnsemblBacteria; AAC77339; AAC77339; b4386.
DR EnsemblBacteria; BAE78375; BAE78375; BAE78375.
DR GeneID; 944865; -.
DR KEGG; ecj:JW4349; -.
DR KEGG; eco:b4386; -.
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; RTCPEDD; -.
DR PhylomeDB; P32099; -.
DR BioCyc; EcoCyc:EG11796-MON; -.
DR BioCyc; MetaCyc:EG11796-MON; -.
DR BRENDA; 6.3.1.20; 2026.
DR UniPathway; UPA00537; UER00594.
DR UniPathway; UPA00537; UER00595.
DR EvolutionaryTrace; P32099; -.
DR PRO; PR:P32099; -.
DR Proteomes; UP000000318; Chromosome.
DR Proteomes; UP000000625; Chromosome.
DR GO; GO:0005737; C:cytoplasm; IBA:GO_Central.
DR GO; GO:0005829; C:cytosol; IDA:EcoCyc.
DR GO; GO:0005524; F:ATP binding; IEA:UniProtKB-KW.
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 Gene3D; 3.30.930.10; -; 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 PANTHER; PTHR12561; PTHR12561; 1.
DR Pfam; PF03099; BPL_LplA_LipB; 1.
DR Pfam; PF10437; Lip_prot_lig_C; 1.
DR SUPFAM; SSF55681; SSF55681; 1.
DR TIGRFAMs; TIGR00545; lipoyltrans; 1.
DR PROSITE; PS51733; BPL_LPL_CATALYTIC; 1.
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 117..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 STRAND 172..175
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
