[1]	NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 2-26, AND CHARACTERIZATION. STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911; PubMed=8206909 [NCBI, ExPASy, EBI, Israel, Japan] Morris T.W., Reed K.E., Cronan J.E. Jr.; "Identification of the gene encoding lipoate-protein ligase A of Escherichia coli. Molecular cloning and characterization of the lplA gene and gene product."; J. Biol. Chem. 269:16091-16100(1994).
[2]	NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. STRAIN=K12 / MG1655 / ATCC 47076; DOI=10.1093/nar/23.12.2105; PubMed=7610040 [NCBI, ExPASy, EBI, Israel, Japan] Burland V.D., Plunkett G. III, Sofia H.J., Daniels D.L., Blattner F.R.; "Analysis of the Escherichia coli genome VI: DNA sequence of the region from 92.8 through 100 minutes."; Nucleic Acids Res. 23:2105-2119(1995).
[3]	NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. STRAIN=K12 / MG1655 / ATCC 47076; DOI=10.1126/science.277.5331.1453; PubMed=9278503 [NCBI, ExPASy, EBI, Israel, Japan] Blattner F.R., Plunkett G. III, Bloch C.A., Perna N.T., Burland V., Riley M., Collado-Vides J., Glasner J.D., Rode C.K., Mayhew G.F., Gregor J., Davis N.W., Kirkpatrick H.A., Goeden M.A., Rose D.J., Mau B., Shao Y.; "The complete genome sequence of Escherichia coli K-12."; Science 277:1453-1474(1997).
[4]	NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911; DOI=10.1038/msb4100049; PubMed=16738553 [NCBI, ExPASy, EBI, Israel, Japan] Hayashi K., Morooka N., Yamamoto Y., Fujita K., Isono K., Choi S., Ohtsubo E., Baba T., Wanner B.L., Mori H., Horiuchi T.; "Highly accurate genome sequences of Escherichia coli K-12 strains MG1655 and W3110."; Mol. Syst. Biol. 2:E1-E5(2006).
[5]	NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 1-68. STRAIN=K12; DOI=10.1016/0378-1119(89)90047-4; PubMed=2684780 [NCBI, ExPASy, EBI, Israel, Japan] Neuwald A.F., Stauffer G.V.; "An Escherichia coli membrane protein with a unique signal sequence."; Gene 82:219-228(1989).
[6]	PARTIAL PROTEIN SEQUENCE, FUNCTION, SUBSTRATE SPECIFICITY, SUBUNIT, AND BIOPHYSICOCHEMICAL PROPERTIES. STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911; PubMed=7639702 [NCBI, ExPASy, EBI, Israel, Japan] Green D.E., Morris T.W., Green J., Cronan J.E. Jr., Guest J.R.; "Purification and properties of the lipoate protein ligase of Escherichia coli."; Biochem. J. 309:853-862(1995).
[7]	VARIANT SER-74. DOI=10.1002/path.1711770102; PubMed=8002607 [NCBI, ExPASy, EBI, Israel, Japan] Morris T.W., Reed K.E., Cronan J.E. Jr.; "Lipoic acid metabolism in Escherichia coli: the lplA and lipB genes define redundant pathways for ligation of lipoyl groups to apoprotein."; J. Bacteriol. 177:1-10(1995).
[8]	CHARACTERIZATION. STRAIN=K12 / JK1; DOI=10.1128/JB.185.5.1582-1589.2003; PubMed=12591875 [NCBI, ExPASy, EBI, Israel, Japan] Jordan S.W., Cronan J.E. Jr.; "The Escherichia coli lipB gene encodes lipoyl (octanoyl)-acyl carrier protein:protein transferase."; J. Bacteriol. 185:1582-1589(2003).
[9]	X-RAY CRYSTALLOGRAPHY (2.4 ANGSTROMS), AND MUTAGENESIS OF SER-73 AND ARG-141. DOI=10.1074/jbc.M505010200; PubMed=16043486 [NCBI, ExPASy, EBI, Israel, Japan] Fujiwara K., Toma S., Okamura-Ikeda K., Motokawa Y., Nakagawa A., Taniguchi H.; "Crystal structure of lipoate-protein ligase A from Escherichia coli. Determination of the lipoic acid-binding site."; J. Biol. Chem. 280:33645-33651(2005).

Comments

FUNCTION: Catalyzes both the ATP-dependent activation of exogenously supplied lipoate to lipoyl-AMP and the transfer of the activated lipoyl onto the lipoyl domains of lipoate-dependent enzymes. Is also able to catalyze very poorly the transfer of lipoyl and octanoyl moiety from their acyl carrier protein.
CATALYTIC ACTIVITY: ATP + lipoate = diphosphate + lipoyl-AMP.
CATALYTIC ACTIVITY: Lipoyl-AMP + protein = protein N⁶-(lipoyl)lysine + AMP.
ENZYME REGULATION: 6-seleno-octanoate, 8-thio-octanoate and 8-seleno-octanoate caused 100%, 50% and 63% inhibition respectively. Inhibited by Cu(2+).

BIOPHYSICOCHEMICAL PROPERTIES:

Kinetic parameters:	K_M=1.9 µM for ATP;
	K_M=1.7 µM for D,L-lipoic acid;
	K_M=152 µM for magnesium ion;
	V_max=40 nmol/min/mg enzyme toward ATP;
	V_max=24 nmol/min/mg enzyme toward D,L-lipoic acid;
pH dependence:	Most active from pH 5.5 to 8.0. Inactive below pH 4.3;

PATHWAY: Protein modification; protein lipoylation via exogenous pathway; protein N(6)-(lipoyl)lysine from lipoic acid: step 1/2 .
PATHWAY: Protein modification; protein lipoylation via exogenous pathway; protein N(6)-(lipoyl)lysine from lipoic acid: step 2/2 .
SUBUNIT: Monomer.
INTERACTION:
P76116:yncE; NbExp=1; IntAct=EBI-553559, EBI-562173;
SUBCELLULAR LOCATION: Cytoplasm.
MISCELLANEOUS: In the transfer reaction, the free carboxyl group of lipoic acid is attached via an amide linkage to the epsilon-amino group of a specific lysine residue of lipoyl domains of lipoate-dependent enzymes.
SIMILARITY: Belongs to the lplA family.

Copyright

Copyrighted by the UniProt Consortium, see http://www.uniprot.org/terms. Distributed under the Creative Commons Attribution-NoDerivs License.

Cross-references

Sequence databases

EMBL

L27665; AAA21740.1; -; Genomic_DNA.	[EMBL / GenBank / DDBJ] [CoDingSequence]
U14003; AAA97282.1; -; Genomic_DNA.	[EMBL / GenBank / DDBJ] [CoDingSequence]
U00096; AAC77339.1; -; Genomic_DNA.	[EMBL / GenBank / DDBJ] [CoDingSequence]
AP009048; BAE78375.1; -; Genomic_DNA.	[EMBL / GenBank / DDBJ] [CoDingSequence]
X03046; CAA26854.1; -; Genomic_DNA.	[EMBL / GenBank / DDBJ] [CoDingSequence]

PIR

A54035; A54035.

RefSeq

AP_004874.1; -.
NP_418803.1; -.

3D structure databases

PDB

1X2G; X-ray; 2.40 A; A/B/C=1-338.	[ExPASy / RCSB / EBI]
1X2H; X-ray; 2.91 A; A/B/C=1-338.	[ExPASy / RCSB / EBI]

Detailed list of linked structures.

PDBsum

1X2G; -.
1X2H; -.

ModBase

P32099.

Protein-protein interaction databases

DIP

DIP:10119N; -.

IntAct

P32099; -.

Enzyme and pathway databases

BioCyc

EcoCyc:EG11796-MON; -.
MetaCyc:EG11796-MON; -.

Organism-specific databases

EchoBASE

EB1744; -.

EcoGene

EG11796; lplA.

Ontologies

GO:0005737;	Cellular component: cytoplasm (inferred from electronic annotation from HAMAP).
GO:0005524;	Molecular function: ATP binding (inferred from electronic annotation from UniProtKB-KW).
GO:0016979;	Molecular function: lipoate-protein ligase activity (inferred from electronic annotation from HAMAP).
GO:0005515;	Molecular function: protein binding (inferred from physical interaction from IntAct).
GO:0016740;	Molecular function: transferase activity (inferred from electronic annotation from InterPro).
GO:0018055;	Biological process: peptidyl-lysine lipoylation (inferred from electronic annotation from HAMAP).
QuickGo view.

Family and domain databases

HAMAP

MF_01602; -; 1.
PBIL [Tree]

InterPro

IPR004143; BPL_LipA_LipB.
IPR005107; CO_DHase_flav_C.
IPR004562; Lipoyltrans.
Graphical view of domain structure.

Gene3D

G3DSA:3.30.390.50; CO_DH_flav_C; 1.

Pfam

PF03099; BPL_LipA_LipB; 1.
Pfam graphical view of domain structure.

TIGRFAMs

TIGR00545; lipoyltrans; 1.

Genome annotation databases

GeneID

944865; -.

GenomeReviews

U00096_GR; b4386.
AP009048_GR; JW4349.

KEGG

ecj:JW4349; -.
eco:b4386; -.

Phylogenomic databases

HOGENOM

P32099; -.

Genome annotation databases

CMR

P32099; b4386.

Other

UniRef

View cluster of proteins with at least 50% / 90% / 100% identity.

Keywords

3D-structure; ATP-binding; Complete proteome; Cytoplasm; Direct protein sequencing; Nucleotide-binding; Transferase.

Features

Feature table viewer

`Key`	`From To`	`Length`	`Description`	`FTId`
`INIT_MET`	`1 1`		`Removed.`
`CHAIN`	`2 338`	`337`	`Lipoate-protein ligase A.`	`PRO_0000209564`
`NP_BIND`	`76 79`	`4`	`ATP (By similarity).`
`BINDING`	`71 71`		`ATP (By similarity).`
`BINDING`	`134 134`		`ATP (By similarity).`
`BINDING`	`134 134`		`Lipoate (By similarity).`
`VARIANT`	`74 74`	`1`	`G -> S (in lplA1 or slr1; selenolipoate resistance mutation).`
`MUTAGEN`	`73 73`		`S->A: 20-fold decrease in affinity for ATP.`
`MUTAGEN`	`141 141`		`R->A: More than 10-fold reduction in Vmax.`
`STRAND`	`3 8`	`6`
`HELIX`	`13 25`	`13`
`STRAND`	`31 37`	`7`
`STRAND`	`40 45`	`6`
`HELIX`	`51 54`	`4`
`HELIX`	`57 63`	`7`
`STRAND`	`66 69`	`4`
`STRAND`	`77 79`	`3`
`STRAND`	`83 91`	`9`
`TURN`	`92 94`	`3`
`HELIX`	`98 110`	`13`
`STRAND`	`115 118`	`4`
`TURN`	`119 121`	`3`
`STRAND`	`122 125`	`4`
`STRAND`	`132 141`	`10`
`STRAND`	`143 155`	`13`
`HELIX`	`160 164`	`5`
`HELIX`	`169 173`	`5`
`HELIX`	`187 189`	`3`
`HELIX`	`196 211`	`16`
`STRAND`	`217 220`	`4`
`HELIX`	`231 238`	`8`
`HELIX`	`241 244`	`4`
`STRAND`	`252 259`	`8`
`STRAND`	`262 271`	`10`
`STRAND`	`274 282`	`9`
`HELIX`	`288 297`	`10`
`STRAND`	`301 303`	`3`
`HELIX`	`304 313`	`10`
`HELIX`	`314 317`	`4`
`HELIX`	`319 321`	`3`
`HELIX`	`322 336`	`15`

Sequence information

Length: 338 AA [This is the length of the unprocessed precursor]

Molecular weight: 37926 Da [This is the MW of the unprocessed precursor]

CRC64: 102788082E182A6F [This is a checksum on the sequence]

        10         20         30         40         50         60 
MSTLRLLISD SYDPWFNLAV EECIFRQMPA TQRVLFLWRN ADTVVIGRAQ NPWKECNTRR 

        70         80         90        100        110        120 
MEEDNVRLAR RSSGGGAVFH DLGNTCFTFM AGKPEYDKTI STSIVLNALN ALGVSAEASG 

       130        140        150        160        170        180 
RNDLVVKTVE GDRKVSGSAY RETKDRGFHH GTLLLNADLS RLANYLNPDK KKLAAKGITS 

       190        200        210        220        230        240 
VRSRVTNLTE LLPGITHEQV CEAITEAFFA HYGERVEAEI ISPNKTPDLP NFAETFARQS 

       250        260        270        280        290        300 
SWEWNFGQAP AFSHLLDERF TWGGVELHFD VEKGHITRAQ VFTDSLNPAP LEALAGRLQG 

       310        320        330 
CLYRADMLQQ ECEALLVDFP EQEKELRELS AWMAGAVR

P32099 in FASTA format

View entry in original UniProtKB/Swiss-Prot format
View entry in raw text format (no links)
Report form for errors/updates in this UniProtKB/Swiss-Prot entry

	BLAST submission on ExPASy/SIB or at NCBI (USA)		Sequence analysis tools: ProtParam, ProtScale, Compute pI/Mw, PeptideMass, PeptideCutter, Dotlet (Java)
	ScanProsite, MotifScan		Submit a homology modeling request to SWISS-MODEL
	NPSA Sequence analysis tools