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MetaCyc Protein: isochorismatase / aryl-carrier protein

Gene: entB Accession Numbers: EG10260 (MetaCyc), b0595, ECK0588

Synonyms: entG

Species: Escherichia coli K-12 substr. MG1655

Subunit composition of isochorismatase / aryl-carrier protein = [EntB]5
         EntB monomer = EntB

Alternative forms of isochorismatase / aryl-carrier protein: EntB isochorismatase / aryl-carrier protein (extended summary available)

EntB is a bifunctional enzyme that is involved in the enterobactin biosynthesis pathway. Enterobactin, a siderophore molecule, is synthesized in response to iron deprivation by formation of an amide bond between 2,3-dihydroxybenzoate (2,3-DHB) and L-serine. EntB is an isochorismatase which catalyzes the synthesis of 2,3-DHB and also serves as an aryl carrier protein (ArCP) in the assembly of enterobactin. [Gehring97]

Apo-EntB has an N-terminal isochorismatase domain and a C-terminal apo-aryl carrier protein (apo-ArCP) domain. The apo-ArCP domain undergoes phosphopantetheinylation catalyzed by phosphopantetheinyl transferase EntD, turning EntB into holo-EntB (EntB isochorismatase / aryl-carrier protein), which serves as a substrate for the 2,3-dihydroxybenzoate-AMP ligase, EntE. [Gehring98, Gehring97]

The surface of the ArCP domain has been mapped and several conserved hydrophobic residues as well as protein interaction surfaces were identified [Lai06a, Lai06b]. The thioesterase, EntH, interacts specifically with the ArCP domain of EntB during enterobactin production [Leduc07].

Based on gel filtration data, EntB was originally suggested to be a pentamer [Rusnak90] and later suggested to be a trimer [Gehring98]. However subsequent gel filtration and crystallographic data showed it to be a dimer [Drake06]. The crystal structure of an engineered fusion protein containing the EntB aryl-carrier protein and the EntE adenylation domain has also been determined to gain insight into the interaction between these two domains [Sundlov12].

EntB is a cytosolic protein which can be released by osmotic shock. Membrane association of EntB has also been demonstrated. [Hantash00]

Overproduction of EntB in a reduced-genome E. coli, promoted biofilm development and maturation [May11].

Citations: [Zhou07, Chen09, Khalil09, Hubrich13]

Locations: cytosol, inner membrane

Map Position: [626,917 -> 627,774]

Molecular Weight of Polypeptide: 32.554 kD (from nucleotide sequence)

pI: 5.33

Unification Links: ASAP:ABE-0002052 , CGSC:821 , EchoBASE:EB0256 , EcoGene:EG10260 , EcoliWiki:b0595 , Mint:MINT-1241343 , ModBase:P0ADI4 , OU-Microarray:b0595 , PortEco:entB , PR:PRO_000022520 , Pride:P0ADI4 , Protein Model Portal:P0ADI4 , RefSeq:NP_415127 , RegulonDB:EG10260 , SMR:P0ADI4 , String:511145.b0595 , Swiss-Model:P0ADI4 , UniProt:P0ADI4

Relationship Links: InterPro:IN-FAMILY:IPR000868 , InterPro:IN-FAMILY:IPR009081 , InterPro:IN-FAMILY:IPR016291 , PDB:Structure:2FQ1 , PDB:Structure:3RG2 , Pfam:IN-FAMILY:PF00550 , Pfam:IN-FAMILY:PF00857 , Prints:IN-FAMILY:PR01398 , Prosite:IN-FAMILY:PS00012 , Prosite:IN-FAMILY:PS50075

Reactions known to consume the compound:

Not in pathways:
isochorismatase / aryl-carrier protein + coenzyme A → EntB isochorismatase / aryl-carrier protein + adenosine 3',5'-bisphosphate

Gene-Reaction Schematic: ?

GO Terms:

Biological Process: GO:0008152 - metabolic process Inferred by computational analysis [GOA01a]
GO:0009239 - enterobactin biosynthetic process Inferred by computational analysis [UniProtGOA12, UniProtGOA11a]
Molecular Function: GO:0005515 - protein binding Inferred from experiment [Leduc07]
GO:0016765 - transferase activity, transferring alkyl or aryl (other than methyl) groups Inferred from experiment [Gehring97]
GO:0042802 - identical protein binding Inferred from experiment [Rajagopala14, Leduc07]
GO:0003824 - catalytic activity Inferred by computational analysis [GOA01a]
GO:0008908 - isochorismatase activity Inferred by computational analysis [GOA01, GOA01a]
GO:0016787 - hydrolase activity Inferred by computational analysis [UniProtGOA11a]
Cellular Component: GO:0005829 - cytosol Inferred from experiment Inferred by computational analysis [DiazMejia09, Ishihama08, Hantash00]
GO:0005886 - plasma membrane Inferred from experiment [Hantash00]

MultiFun Terms: cell structure membrane
information transfer protein related Non-ribosomal peptide synthetase
metabolism biosynthesis of building blocks cofactors, small molecule carriers enterochelin (enterobactin)

Imported from EcoCyc 04-Feb-2014 by Paley S , SRI International

Sequence Features

Feature Class Location Citations Comment
Protein-Segment 26 -> 208
UniProt: Isochorismate lyase; Sequence Annotation Type: region of interest;
Conserved-Region 214 -> 285
UniProt: Acyl carrier;
Modified-Residue 245
UniProt: O-(pantetheine 4'-phosphoryl)serine; Non-Experimental Qualifier: potential;

10/20/97 Gene b0595 from Blattner lab Genbank (v. M52) entry merged into EcoCyc gene EG10260; confirmed by SwissProt match.


Chen09: Chen D, Wu R, Bryan TL, Dunaway-Mariano D (2009). "In vitro kinetic analysis of substrate specificity in enterobactin biosynthetic lower pathway enzymes provides insight into the biochemical function of the hot dog-fold thioesterase EntH." Biochemistry 48(3);511-3. PMID: 19119850

DiazMejia09: Diaz-Mejia JJ, Babu M, Emili A (2009). "Computational and experimental approaches to chart the Escherichia coli cell-envelope-associated proteome and interactome." FEMS Microbiol Rev 33(1);66-97. PMID: 19054114

Drake06: Drake EJ, Nicolai DA, Gulick AM (2006). "Structure of the EntB multidomain nonribosomal peptide synthetase and functional analysis of its interaction with the EntE adenylation domain." Chem Biol 13(4);409-19. PMID: 16632253

Gehring97: Gehring AM, Bradley KA, Walsh CT (1997). "Enterobactin biosynthesis in Escherichia coli: isochorismate lyase (EntB) is a bifunctional enzyme that is phosphopantetheinylated by EntD and then acylated by EntE using ATP and 2,3-dihydroxybenzoate." Biochemistry 1997;36(28);8495-503. PMID: 9214294

Gehring98: Gehring AM, Mori I, Walsh CT (1998). "Reconstitution and characterization of the Escherichia coli enterobactin synthetase from EntB, EntE, and EntF." Biochemistry 1998;37(8);2648-59. PMID: 9485415

GOA01: GOA, MGI (2001). "Gene Ontology annotation based on Enzyme Commission mapping." Genomics 74;121-128.

GOA01a: GOA, DDB, FB, MGI, ZFIN (2001). "Gene Ontology annotation through association of InterPro records with GO terms."

Hantash00: Hantash FM, Earhart CF (2000). "Membrane association of the Escherichia coli enterobactin synthase proteins EntB/G, EntE, and EntF." J Bacteriol 182(6);1768-73. PMID: 10692387

Hubrich13: Hubrich F, Mordhorst S, Andexer JN (2013). "Cinnamic acid derivatives as inhibitors for chorismatases and isochorismatases." Bioorg Med Chem Lett 23(5);1477-81. PMID: 23380376

Ishihama08: Ishihama Y, Schmidt T, Rappsilber J, Mann M, Hartl FU, Kerner MJ, Frishman D (2008). "Protein abundance profiling of the Escherichia coli cytosol." BMC Genomics 9;102. PMID: 18304323

Khalil09: Khalil S, Pawelek PD (2009). "Ligand-induced conformational rearrangements promote interaction between the Escherichia coli enterobactin biosynthetic proteins EntE and EntB." J Mol Biol 393(3);658-71. PMID: 19699210

Lai06a: Lai JR, Fischbach MA, Liu DR, Walsh CT (2006). "A protein interaction surface in nonribosomal peptide synthesis mapped by combinatorial mutagenesis and selection." Proc Natl Acad Sci U S A 103(14);5314-9. PMID: 16567620

Lai06b: Lai JR, Fischbach MA, Liu DR, Walsh CT (2006). "Localized protein interaction surfaces on the EntB carrier protein revealed by combinatorial mutagenesis and selection." J Am Chem Soc 128(34);11002-3. PMID: 16925399

Leduc07: Leduc D, Battesti A, Bouveret E (2007). "The hotdog thioesterase EntH (YbdB) plays a role in vivo in optimal enterobactin biosynthesis by interacting with the ArCP domain of EntB." J Bacteriol 189(19);7112-26. PMID: 17675380

May11: May T, Okabe S (2011). "Enterobactin is required for biofilm development in reduced-genome Escherichia coli." Environ Microbiol 13(12);3149-62. PMID: 21980953

Rajagopala14: Rajagopala SV, Sikorski P, Kumar A, Mosca R, Vlasblom J, Arnold R, Franca-Koh J, Pakala SB, Phanse S, Ceol A, Hauser R, Siszler G, Wuchty S, Emili A, Babu M, Aloy P, Pieper R, Uetz P (2014). "The binary protein-protein interaction landscape of Escherichia coli." Nat Biotechnol 32(3);285-90. PMID: 24561554

Rusnak90: Rusnak F, Liu J, Quinn N, Berchtold GA, Walsh CT (1990). "Subcloning of the enterobactin biosynthetic gene entB: expression, purification, characterization, and substrate specificity of isochorismatase." Biochemistry 1990;29(6);1425-35. PMID: 2139796

Sundlov12: Sundlov JA, Shi C, Wilson DJ, Aldrich CC, Gulick AM (2012). "Structural and functional investigation of the intermolecular interaction between NRPS adenylation and carrier protein domains." Chem Biol 19(2);188-98. PMID: 22365602

UniProt09: UniProt Consortium (2009). "UniProt version 15.8 released on 2009-10-01 00:00:00." Database.

UniProt10a: UniProt Consortium (2010). "UniProt version 2010-07 released on 2010-06-15 00:00:00." Database.

UniProtGOA11a: UniProt-GOA (2011). "Gene Ontology annotation based on manual assignment of UniProtKB keywords in UniProtKB/Swiss-Prot entries."

UniProtGOA12: UniProt-GOA (2012). "Gene Ontology annotation based on UniPathway vocabulary mapping."

Zhou07: Zhou Z, Lai JR, Walsh CT (2007). "Directed evolution of aryl carrier proteins in the enterobactin synthetase." Proc Natl Acad Sci U S A 104(28);11621-6. PMID: 17606920

Report Errors or Provide Feedback
Please cite the following article in publications resulting from the use of MetaCyc: Caspi et al, Nucleic Acids Research 42:D459-D471 2014
Page generated by SRI International Pathway Tools version 18.5 on Wed Mar 4, 2015, biocyc11.