P11454 · ENTF_ECOLI
- ProteinEnterobactin synthase component F
- GeneentF
- StatusUniProtKB reviewed (Swiss-Prot)
- Organism
- Amino acids1293 (go to sequence)
- Protein existenceEvidence at protein level
- Annotation score5/5
Function
function
Involved in the biosynthesis of the siderophore enterobactin (enterochelin), which is a macrocyclic trimeric lactone of N-(2,3-dihydroxybenzoyl)-serine (PubMed:1338974, PubMed:1826089, PubMed:216414, PubMed:9485415).
EntF catalyzes the activation of L-serine via ATP-dependent PPi exchange reaction to form seryladenylate (PubMed:10688898, PubMed:1338974, PubMed:1826089, PubMed:9485415).
Activated L-serine is loaded onto the peptidyl carrier domain via a thioester linkage to the phosphopanthetheine moiety, forming seryl-S-Ppant-EntF (PubMed:9485415).
EntF acts then as the sole catalyst for the formation of the three amide and three ester linkages found in enterobactin, using seryladenylate and 2,3-dihydroxybenzoate-S-Ppant-EntB (DHB-S-Ppant-EntB) as substrates, via the formation of a DHB-Ser-S-Ppant-EntF intermediate (PubMed:9485415).
EntF catalyzes the activation of L-serine via ATP-dependent PPi exchange reaction to form seryladenylate (PubMed:10688898, PubMed:1338974, PubMed:1826089, PubMed:9485415).
Activated L-serine is loaded onto the peptidyl carrier domain via a thioester linkage to the phosphopanthetheine moiety, forming seryl-S-Ppant-EntF (PubMed:9485415).
EntF acts then as the sole catalyst for the formation of the three amide and three ester linkages found in enterobactin, using seryladenylate and 2,3-dihydroxybenzoate-S-Ppant-EntB (DHB-S-Ppant-EntB) as substrates, via the formation of a DHB-Ser-S-Ppant-EntF intermediate (PubMed:9485415).
Catalytic activity
- 3 2,3-dihydroxybenzoate + 6 ATP + 3 L-serine = 6 AMP + 6 diphosphate + enterobactin + 4 H+
Cofactor
Note: Binds 1 phosphopantetheine covalently.
Activity regulation
Adenylation activity is increased in the presence of the MbtH-like protein YbdZ.
Kinetics
KM | SUBSTRATE | pH | TEMPERATURE[C] | NOTES | EVIDENCE | |
---|---|---|---|---|---|---|
0.26 mM | L-serine | |||||
0.75 mM | ATP | |||||
5 mM | O-acetyl-L-serine | |||||
8 mM | L-beta-chloroalanine | |||||
3.1 mM | S-methyl-L-cysteine |
kcat is 760 min-1 with L-serine as substrate. kcat is 660 min-1 with ATP as substrate. kcat is 720 min-1 with O-acetyl-L-serine as substrate. kcat is 870 min-1 with L-beta-chloroalanine as substrate. kcat is 38 min-1 with S-methyl-L-cysteine as substrate.
Pathway
Siderophore biosynthesis; enterobactin biosynthesis.
Features
Showing features for active site.
Type | ID | Position(s) | Description | |||
---|---|---|---|---|---|---|
Active site | 1271 | Proton acceptor; for thioesterase activity | ||||
Sequence: H |
GO annotations
Aspect | Term | |
---|---|---|
Cellular Component | cytoplasm | |
Cellular Component | cytosol | |
Cellular Component | enterobactin synthetase complex | |
Cellular Component | plasma membrane | |
Molecular Function | 2,3-dihydroxybenzoate-serine ligase activity | |
Molecular Function | ATP binding | |
Molecular Function | nucleotidyltransferase activity | |
Molecular Function | phosphopantetheine binding | |
Biological Process | amino acid activation for nonribosomal peptide biosynthetic process | |
Biological Process | enterobactin biosynthetic process |
Keywords
- Molecular function
- Biological process
- Ligand
Enzyme and pathway databases
Protein family/group databases
Names & Taxonomy
Protein names
- Recommended nameEnterobactin synthase component F
- EC number
- Alternative names
Including 1 domain:
- Recommended nameL-serine--[L-seryl-carrier protein] ligase
- EC number
- Alternative names
Gene names
Organism names
- Organism
- Strains
- Taxonomic lineageBacteria > Pseudomonadota > Gammaproteobacteria > Enterobacterales > Enterobacteriaceae > Escherichia
Accessions
- Primary accessionP11454
- Secondary accessions
Proteomes
Subcellular Location
Phenotypes & Variants
Features
Showing features for mutagenesis.
Type | ID | Position(s) | Description | |||
---|---|---|---|---|---|---|
Mutagenesis | 1077 | Retains 2% of activity. | ||||
Sequence: F → A | ||||||
Mutagenesis | 1079 | Abolishes enterobactin production. | ||||
Sequence: W → A | ||||||
Mutagenesis | 1080 | Retains 50% of activity. | ||||
Sequence: Q → A | ||||||
Mutagenesis | 1138 | No enterobactin synthase activity. This mutant holo-EntF is still able to adenylate serine and to autoacylate itself with serine. | ||||
Sequence: S → A | ||||||
Mutagenesis | 1138 | 1000-fold decrease in kcat. Releases both enterobactin an linear dimers (DHB-Ser)2. | ||||
Sequence: S → C | ||||||
Mutagenesis | 1165 | 20-fold decrease in kcat. | ||||
Sequence: D → A | ||||||
Mutagenesis | 1165 | 200-fold decrease in kcat. | ||||
Sequence: D → S | ||||||
Mutagenesis | 1271 | 10000-fold decrease in kcat. Releases only enterobactin, but accumulates both DHB-Ser-O-TE and (DHB-Ser)2-O-TE acyl enzyme intermediates. | ||||
Sequence: H → A |
PTM/Processing
Features
Showing features for chain, modified residue.
Type | ID | Position(s) | Description | |||
---|---|---|---|---|---|---|
Chain | PRO_0000193077 | 1-1293 | Enterobactin synthase component F | |||
Sequence: MSQHLPLVAAQPGIWMAEKLSELPSAWSVAHYVELTGEVDSPLLARAVVAGLAQADTLRMRFTEDNGEVWQWVDDALTFELPEIIDLRTNIDPHGTAQALMQADLQQDLRVDSGKPLVFHQLIQVADNRWYWYQRYHHLLVDGFSFPAITRQIANIYCTWLRGEPTPASPFTPFADVVEEYQQYRESEAWQRDAAFWAEQRRQLPPPASLSPAPLPGRSASADILRLKLEFTDGEFRQLATQLSGVQRTDLALALAALWLGRLCNRMDYAAGFIFMRRLGSAALTATGPVLNVLPLGIHIAAQETLPELATRLAAQLKKMRRHQRYDAEQIVRDSGRAAGDEPLFGPVLNIKVFDYQLDIPDVQAQTHTLATGPVNDLELALFPDVHGDLSIEILANKQRYDEPTLIQHAERLKMLIAQFAADPALLCGDVDIMLPGEYAQLAQLNATQVEIPETTLSALVAEQAAKTPDAPALADARYLFSYREMREQVVALANLLRERGVKPGDSVAVALPRSVFLTLALHAIVEAGAAWLPLDTGYPDDRLKMMLEDARPSLLITTDDQLPRFSDVPNLTSLCYNAPLTPQGSAPLQLSQPHHTAYIIFTSGSTGRPKGVMVGQTAIVNRLLWMQNHYPLTGEDVVAQKTPCSFDVSVWEFFWPFIAGAKLVMAEPEAHRDPLAMQQFFAEYGVTTTHFVPSMLAAFVASLTPQTARQSCATLKQVFCSGEALPADLCREWQQLTGAPLHNLYGPTEAAVDVSWYPAFGEELAQVRGSSVPIGYPVWNTGLRILDAMMHPVPPGVAGDLYLTGIQLAQGYLGRPDLTASRFIADPFAPGERMYRTGDVARWLDNGAVEYLGRSDDQLKIRGQRIELGEIDRVMQALPDVEQAVTHACVINQAAATGGDARQLVGYLVSQSGLPLDTSALQAQLRETLPPHMVPVVLLQLPQLPLSANGKLDRKALPLPELKAQAPGRAPKAGSETIIAAAFSSLLGCDVQDADADFFALGGHSLLAMKLAAQLSRQVARQVTPGQVMVASTVAKLATIIDAEEDSTRRMGFETILPLREGNGPTLFCFHPASGFAWQFSVLSRYLDPQWSIIGIQSPRPNGPMQTAANLDEVCEAHLATLLEQQPHGPYYLLGYSLGGTLAQGIAARLRARGEQVAFLGLLDTWPPETQNWQEKEANGLDPEVLAEINREREAFLAAQQGSTSTELFTTIEGNYADAVRLLTTAHSVPFDGKATLFVAERTLQEGMSPERAWSPWIAELDIYRQDCAHVDIISPGTFEKIGPIIRATLNR | ||||||
Modified residue | 1006 | O-(pantetheine 4'-phosphoryl)serine | ||||
Sequence: S |
Post-translational modification
4'-phosphopantetheine is transferred from CoA to a specific serine of apo-EntF by EntD (PubMed:9485415).
Holo-EntF so formed is then acylated with seryl-AMP (PubMed:9485415).
Holo-EntF so formed is then acylated with seryl-AMP (PubMed:9485415).
Keywords
- PTM
Proteomic databases
Expression
Induction
Transcriptionally regulated by iron and the fur protein.
Interaction
Subunit
Proteins EntB, EntD, EntE and EntF are the component of the enterobactin synthase (PubMed:216414, PubMed:9485415).
Components probably do not form a stable complex (PubMed:9485415).
EntF acts as a catalytic monomer (PubMed:9485415).
Interacts with the MbtH-like protein YbdZ. YbdZ binds to the adenylation domain, but does not alter the structure of the domain (PubMed:27597544).
Components probably do not form a stable complex (PubMed:9485415).
EntF acts as a catalytic monomer (PubMed:9485415).
Interacts with the MbtH-like protein YbdZ. YbdZ binds to the adenylation domain, but does not alter the structure of the domain (PubMed:27597544).
Protein-protein interaction databases
Structure
Family & Domains
Features
Showing features for region, domain.
Type | ID | Position(s) | Description | |||
---|---|---|---|---|---|---|
Region | 1-301 | Elongation/condensation | ||||
Sequence: MSQHLPLVAAQPGIWMAEKLSELPSAWSVAHYVELTGEVDSPLLARAVVAGLAQADTLRMRFTEDNGEVWQWVDDALTFELPEIIDLRTNIDPHGTAQALMQADLQQDLRVDSGKPLVFHQLIQVADNRWYWYQRYHHLLVDGFSFPAITRQIANIYCTWLRGEPTPASPFTPFADVVEEYQQYRESEAWQRDAAFWAEQRRQLPPPASLSPAPLPGRSASADILRLKLEFTDGEFRQLATQLSGVQRTDLALALAALWLGRLCNRMDYAAGFIFMRRLGSAALTATGPVLNVLPLGIHIA | ||||||
Region | 482-887 | Adenylatione | ||||
Sequence: SYREMREQVVALANLLRERGVKPGDSVAVALPRSVFLTLALHAIVEAGAAWLPLDTGYPDDRLKMMLEDARPSLLITTDDQLPRFSDVPNLTSLCYNAPLTPQGSAPLQLSQPHHTAYIIFTSGSTGRPKGVMVGQTAIVNRLLWMQNHYPLTGEDVVAQKTPCSFDVSVWEFFWPFIAGAKLVMAEPEAHRDPLAMQQFFAEYGVTTTHFVPSMLAAFVASLTPQTARQSCATLKQVFCSGEALPADLCREWQQLTGAPLHNLYGPTEAAVDVSWYPAFGEELAQVRGSSVPIGYPVWNTGLRILDAMMHPVPPGVAGDLYLTGIQLAQGYLGRPDLTASRFIADPFAPGERMYRTGDVARWLDNGAVEYLGRSDDQLKIRGQRIELGEIDRVMQALPDVEQAVT | ||||||
Domain | 971-1046 | Carrier | ||||
Sequence: APKAGSETIIAAAFSSLLGCDVQDADADFFALGGHSLLAMKLAAQLSRQVARQVTPGQVMVASTVAKLATIIDAEE | ||||||
Region | 1066-1293 | Thioesterase | ||||
Sequence: PTLFCFHPASGFAWQFSVLSRYLDPQWSIIGIQSPRPNGPMQTAANLDEVCEAHLATLLEQQPHGPYYLLGYSLGGTLAQGIAARLRARGEQVAFLGLLDTWPPETQNWQEKEANGLDPEVLAEINREREAFLAAQQGSTSTELFTTIEGNYADAVRLLTTAHSVPFDGKATLFVAERTLQEGMSPERAWSPWIAELDIYRQDCAHVDIISPGTFEKIGPIIRATLNR |
Domain
Modular protein that contains an elongation/condensation domain, an adenylation domain which activates the serine residue into an aminoacyl-AMP ester, a peptidyl carrier protein domain which bears a phosphopantetheinyl arm to attach the activated amino acid and a thioesterase domain (Probable). The thioesterase domain is both a cyclotrimerizing lactone synthetase and an elongation catalyst for ester-bond formation between covalently tethered DHB-Ser moieties (PubMed:10375542).
The carrier-thioesterase di-domain forms a compact structure with a well-defined domain interface; the two active sites are at a suitable distance for substrate transfer from the peptidyl carrier protein domain to the thioesterase domain (PubMed:18704088).
Phosphopantetheinylation leads to enhanced interaction between the two domains (PubMed:22118682).
The carrier-thioesterase di-domain forms a compact structure with a well-defined domain interface; the two active sites are at a suitable distance for substrate transfer from the peptidyl carrier protein domain to the thioesterase domain (PubMed:18704088).
Phosphopantetheinylation leads to enhanced interaction between the two domains (PubMed:22118682).
Sequence similarities
Belongs to the ATP-dependent AMP-binding enzyme family. EntF subfamily.
Phylogenomic databases
Family and domain databases
Sequence
- Sequence statusComplete
- Length1,293
- Mass (Da)141,991
- Last updated1997-11-01 v3
- ChecksumF3FCBD3836A39358
Sequence caution
Features
Showing features for sequence conflict.
Type | ID | Position(s) | Description | |||
---|---|---|---|---|---|---|
Sequence conflict | 441-442 | in Ref. 1; AAA92015 | ||||
Sequence: QL → HV |
Keywords
- Technical term
Sequence databases
Nucleotide Sequence | Protein Sequence | Molecule Type | Status | |
---|---|---|---|---|
M60177 EMBL· GenBank· DDBJ | AAA92015.1 EMBL· GenBank· DDBJ | Genomic DNA | ||
U82598 EMBL· GenBank· DDBJ | AAB40785.1 EMBL· GenBank· DDBJ | Genomic DNA | Different initiation | |
U00096 EMBL· GenBank· DDBJ | AAC73687.1 EMBL· GenBank· DDBJ | Genomic DNA | ||
AP009048 EMBL· GenBank· DDBJ | BAE76341.1 EMBL· GenBank· DDBJ | Genomic DNA | ||
M17354 EMBL· GenBank· DDBJ | AAA23727.1 EMBL· GenBank· DDBJ | Genomic DNA | ||
J04216 EMBL· GenBank· DDBJ | AAA23759.1 EMBL· GenBank· DDBJ | Genomic DNA |