P11454 · ENTF_ECOLI

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).

Catalytic activity

Cofactor

pantetheine 4'-phosphate (UniProtKB | Rhea| CHEBI:47942 )

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 mML-serine
0.75 mMATP
5 mMO-acetyl-L-serine
8 mML-beta-chloroalanine
3.1 mMS-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.

TypeIDPosition(s)Description
Active site1271Proton acceptor; for thioesterase activity

GO annotations

AspectTerm
Cellular Componentcytoplasm
Cellular Componentcytosol
Cellular Componententerobactin synthetase complex
Cellular Componentplasma membrane
Molecular Function2,3-dihydroxybenzoate-serine ligase activity
Molecular FunctionATP binding
Molecular Functionnucleotidyltransferase activity
Molecular Functionphosphopantetheine binding
Biological Processamino acid activation for nonribosomal peptide biosynthetic process
Biological Processenterobactin biosynthetic process

Keywords

Enzyme and pathway databases

Protein family/group databases

Names & Taxonomy

Protein names

  • Recommended name
    Enterobactin synthase component F
  • EC number
  • Alternative names
    • Enterochelin synthase F
    • Nonribosomal peptide synthetase EntF

Including 1 domain:

  • Recommended name
    L-serine--[L-seryl-carrier protein] ligase
  • EC number
  • Alternative names
    • Serine-activating enzyme
    • Seryl-AMP ligase

Gene names

    • Name
      entF
    • Ordered locus names
      b0586, JW0578

Organism names

  • Taxonomic identifier
  • Strains
    • K12
    • K12 / MG1655 / ATCC 47076
    • K12 / W3110 / ATCC 27325 / DSM 5911
    • K12 / MC4100 / ATCC 35695 / DSM 6574
  • Taxonomic lineage
    Bacteria > Pseudomonadota > Gammaproteobacteria > Enterobacterales > Enterobacteriaceae > Escherichia

Accessions

  • Primary accession
    P11454
  • Secondary accessions
    • P75723
    • P77095
    • Q2MBL5

Proteomes

Subcellular Location

Keywords

Phenotypes & Variants

Features

Showing features for mutagenesis.

TypeIDPosition(s)Description
Mutagenesis1077Retains 2% of activity.
Mutagenesis1079Abolishes enterobactin production.
Mutagenesis1080Retains 50% of activity.
Mutagenesis1138No enterobactin synthase activity. This mutant holo-EntF is still able to adenylate serine and to autoacylate itself with serine.
Mutagenesis11381000-fold decrease in kcat. Releases both enterobactin an linear dimers (DHB-Ser)2.
Mutagenesis116520-fold decrease in kcat.
Mutagenesis1165200-fold decrease in kcat.
Mutagenesis127110000-fold decrease in kcat. Releases only enterobactin, but accumulates both DHB-Ser-O-TE and (DHB-Ser)2-O-TE acyl enzyme intermediates.

PTM/Processing

Features

Showing features for chain, modified residue.

TypeIDPosition(s)Description
ChainPRO_00001930771-1293Enterobactin synthase component F
Modified residue1006O-(pantetheine 4'-phosphoryl)serine

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).

Keywords

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).

Protein-protein interaction databases

Family & Domains

Features

Showing features for region, domain.

TypeIDPosition(s)Description
Region1-301Elongation/condensation
Region482-887Adenylatione
Domain971-1046Carrier
Region1066-1293Thioesterase

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).

Sequence similarities

Phylogenomic databases

Family and domain databases

Sequence

  • Sequence status
    Complete
  • Length
    1,293
  • Mass (Da)
    141,991
  • Last updated
    1997-11-01 v3
  • Checksum
    F3FCBD3836A39358
MSQHLPLVAAQPGIWMAEKLSELPSAWSVAHYVELTGEVDSPLLARAVVAGLAQADTLRMRFTEDNGEVWQWVDDALTFELPEIIDLRTNIDPHGTAQALMQADLQQDLRVDSGKPLVFHQLIQVADNRWYWYQRYHHLLVDGFSFPAITRQIANIYCTWLRGEPTPASPFTPFADVVEEYQQYRESEAWQRDAAFWAEQRRQLPPPASLSPAPLPGRSASADILRLKLEFTDGEFRQLATQLSGVQRTDLALALAALWLGRLCNRMDYAAGFIFMRRLGSAALTATGPVLNVLPLGIHIAAQETLPELATRLAAQLKKMRRHQRYDAEQIVRDSGRAAGDEPLFGPVLNIKVFDYQLDIPDVQAQTHTLATGPVNDLELALFPDVHGDLSIEILANKQRYDEPTLIQHAERLKMLIAQFAADPALLCGDVDIMLPGEYAQLAQLNATQVEIPETTLSALVAEQAAKTPDAPALADARYLFSYREMREQVVALANLLRERGVKPGDSVAVALPRSVFLTLALHAIVEAGAAWLPLDTGYPDDRLKMMLEDARPSLLITTDDQLPRFSDVPNLTSLCYNAPLTPQGSAPLQLSQPHHTAYIIFTSGSTGRPKGVMVGQTAIVNRLLWMQNHYPLTGEDVVAQKTPCSFDVSVWEFFWPFIAGAKLVMAEPEAHRDPLAMQQFFAEYGVTTTHFVPSMLAAFVASLTPQTARQSCATLKQVFCSGEALPADLCREWQQLTGAPLHNLYGPTEAAVDVSWYPAFGEELAQVRGSSVPIGYPVWNTGLRILDAMMHPVPPGVAGDLYLTGIQLAQGYLGRPDLTASRFIADPFAPGERMYRTGDVARWLDNGAVEYLGRSDDQLKIRGQRIELGEIDRVMQALPDVEQAVTHACVINQAAATGGDARQLVGYLVSQSGLPLDTSALQAQLRETLPPHMVPVVLLQLPQLPLSANGKLDRKALPLPELKAQAPGRAPKAGSETIIAAAFSSLLGCDVQDADADFFALGGHSLLAMKLAAQLSRQVARQVTPGQVMVASTVAKLATIIDAEEDSTRRMGFETILPLREGNGPTLFCFHPASGFAWQFSVLSRYLDPQWSIIGIQSPRPNGPMQTAANLDEVCEAHLATLLEQQPHGPYYLLGYSLGGTLAQGIAARLRARGEQVAFLGLLDTWPPETQNWQEKEANGLDPEVLAEINREREAFLAAQQGSTSTELFTTIEGNYADAVRLLTTAHSVPFDGKATLFVAERTLQEGMSPERAWSPWIAELDIYRQDCAHVDIISPGTFEKIGPIIRATLNR

Sequence caution

The sequence AAB40785.1 differs from that shown. Reason: Erroneous initiation Extended N-terminus.

Features

Showing features for sequence conflict.

TypeIDPosition(s)Description
Sequence conflict441-442in Ref. 1; AAA92015

Keywords

Sequence databases

Nucleotide SequenceProtein SequenceMolecule TypeStatus
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

Genome annotation databases

Similar Proteins

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