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Escherichia coli K-12 substr. MG1655 Polypeptide: apo-serine activating enzyme



Gene: entF Accession Numbers: EG10264 (EcoCyc), b0586, ECK0579

Regulation Summary Diagram: ?

Alternative forms of apo-serine activating enzyme: aryl carrier protein / L-seryl-AMP synthase (extended summary available)

Summary:
EntF is part of 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. EntF is an activating enzyme for L-serine and catalyzes the formation of an L-seryl-AMP-EntF complex. It activates and binds Ser to itself via a thioester linkage to bound 4'-phosphopantetheine. [Rusnak91, Gehring98, Pettis87, Reichert92]

EntF is one of three proteins (also EntE and EntB) which comprises the non ribosomal peptide synthetase which is the E. coli enterobactin synthetase system. EntF contains the complete serine module and a C-terminal domain that catalyzes the formation of the trilactone ring. [Gehring97, Gehring98]. The apo form of EntF serves as a substrate for EntD [Gehring98].

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

Citations: [Ehmann00, Zhou07]

Gene Citations: [Hunt94]

Locations: inner membrane, cytosol

Map Position: [613,380 -> 617,261] (13.22 centisomes)
Length: 3882 bp / 1293 aa

Molecular Weight of Polypeptide: 141.99 kD (from nucleotide sequence), 142.0 kD (experimental) [Reichert92 ]

pI: 5.3

Unification Links: ASAP:ABE-0002021 , CGSC:817 , DIP:DIP-9516N , EchoBASE:EB0260 , EcoGene:EG10264 , EcoliWiki:b0586 , ModBase:P11454 , OU-Microarray:b0586 , PortEco:entF , PR:PRO_000022524 , Pride:P11454 , Protein Model Portal:P11454 , RegulonDB:EG10264 , SMR:P11454 , String:511145.b0586 , UniProt:P11454

Relationship Links: InterPro:IN-FAMILY:IPR000873 , InterPro:IN-FAMILY:IPR001031 , InterPro:IN-FAMILY:IPR001242 , InterPro:IN-FAMILY:IPR006162 , InterPro:IN-FAMILY:IPR009081 , InterPro:IN-FAMILY:IPR010071 , InterPro:IN-FAMILY:IPR020845 , InterPro:IN-FAMILY:IPR025110 , PDB:Structure:2ROQ , PDB:Structure:3TEJ , Pfam:IN-FAMILY:PF00501 , Pfam:IN-FAMILY:PF00550 , Pfam:IN-FAMILY:PF00668 , Pfam:IN-FAMILY:PF00975 , Pfam:IN-FAMILY:PF13193 , Prosite:IN-FAMILY:PS00012 , Prosite:IN-FAMILY:PS00455 , Prosite:IN-FAMILY:PS50075

In Paralogous Gene Group: 13 (9 members)

Reactions known to consume the compound:

Not in pathways:
apo-serine activating enzyme + coenzyme A → adenosine 3',5'-bisphosphate + aryl carrier protein / L-seryl-AMP synthase

Gene-Reaction Schematic: ?

Genetic Regulation Schematic: ?

GO Terms:

Biological Process: GO:0009239 - enterobactin biosynthetic process Inferred from experiment Inferred by computational analysis [UniProtGOA12, UniProtGOA11a, Gehring98]
GO:0043041 - amino acid activation for nonribosomal peptide biosynthetic process Inferred from experiment [Rusnak91]
GO:0008152 - metabolic process Inferred by computational analysis [UniProtGOA11a, GOA01]
GO:0009058 - biosynthetic process Inferred by computational analysis [GOA01]
Molecular Function: GO:0031177 - phosphopantetheine binding Inferred from experiment [Lambalot96]
GO:0000166 - nucleotide binding Inferred by computational analysis [UniProtGOA11a]
GO:0003824 - catalytic activity Inferred by computational analysis [UniProtGOA11a, GOA01]
GO:0005524 - ATP binding Inferred by computational analysis [UniProtGOA11a]
GO:0016740 - transferase activity Inferred by computational analysis [UniProtGOA11a]
GO:0016788 - hydrolase activity, acting on ester bonds Inferred by computational analysis [GOA01]
GO:0016874 - ligase activity Inferred by computational analysis [UniProtGOA11a]
Cellular Component: GO:0005829 - cytosol Inferred from experiment [Ishihama08, Hantash00]
GO:0005886 - plasma membrane Inferred from experiment [Hantash00]
GO:0009366 - enterobactin synthetase complex Inferred from experiment [Gehring98]

MultiFun Terms: cell processes adaptations Fe aquisition
cell structure membrane
information transfer protein related Non-ribosomal peptide synthetase
metabolism biosynthesis of building blocks cofactors, small molecule carriers Coenzyme A and its modification
metabolism biosynthesis of building blocks cofactors, small molecule carriers enterochelin (enterobactin)

Essentiality data for entF knockouts: ?

Growth Medium Growth? T (°C) O2 pH Osm/L Growth Observations
LB enriched Yes 37 Aerobic 6.95   Yes [Gerdes03, Comment 1]
LB Lennox Yes 37 Aerobic 7   Yes [Baba06, Comment 2]
M9 medium with 1% glycerol Yes 37 Aerobic 7.2 0.35 Yes [Joyce06, Comment 3]
MOPS medium with 0.4% glucose Yes 37 Aerobic 7.2 0.22 Yes [Baba06, Comment 2]
Yes [Feist07, Comment 4]

Sequence Features

Feature Class Location Citations Comment
Protein-Segment 1 -> 301
[UniProt10a]
UniProt: Elongation/condensation; Sequence Annotation Type: region of interest;
Sequence-Conflict 441 -> 442
[Rusnak91, UniProt10]
Alternate sequence: QL → HV; UniProt: (in Ref. 1; AAA92015);
Protein-Segment 482 -> 887
[UniProt10a]
UniProt: Adenylation; Sequence Annotation Type: region of interest;
Conserved-Region 976 -> 1043
[UniProt09]
UniProt: Acyl carrier;
Modified-Residue 1006
[UniProt10a]
UniProt: O-(pantetheine 4'-phosphoryl)serine; Non-Experimental Qualifier: probable;
Protein-Segment 1066 -> 1293
[UniProt10a]
UniProt: Thioesterase; Sequence Annotation Type: region of interest;
Mutagenesis-Variant 1138
[Gehring98, UniProt11]
Alternate sequence: S → A; UniProt: No enterobactin synthase activity. This mutant holo-EntF is still able to adenylate serine and to autoacylate itself with serine.


Gene Local Context (not to scale): ?

Transcription Units:

Notes:

History:
10/20/97 Gene b0586 from Blattner lab Genbank (v. M52) entry merged into EcoCyc gene EG10264; confirmed by SwissProt match.


References

Baba06: Baba T, Ara T, Hasegawa M, Takai Y, Okumura Y, Baba M, Datsenko KA, Tomita M, Wanner BL, Mori H (2006). "Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection." Mol Syst Biol 2;2006.0008. PMID: 16738554

Ehmann00: Ehmann DE, Shaw-Reid CA, Losey HC, Walsh CT (2000). "The EntF and EntE adenylation domains of Escherichia coli enterobactin synthetase: sequestration and selectivity in acyl-AMP transfers to thiolation domain cosubstrates." Proc Natl Acad Sci U S A 97(6);2509-14. PMID: 10688898

Feist07: Feist AM, Henry CS, Reed JL, Krummenacker M, Joyce AR, Karp PD, Broadbelt LJ, Hatzimanikatis V, Palsson BO (2007). "A genome-scale metabolic reconstruction for Escherichia coli K-12 MG1655 that accounts for 1260 ORFs and thermodynamic information." Mol Syst Biol 3;121. PMID: 17593909

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

Gerdes03: Gerdes SY, Scholle MD, Campbell JW, Balazsi G, Ravasz E, Daugherty MD, Somera AL, Kyrpides NC, Anderson I, Gelfand MS, Bhattacharya A, Kapatral V, D'Souza M, Baev MV, Grechkin Y, Mseeh F, Fonstein MY, Overbeek R, Barabasi AL, Oltvai ZN, Osterman AL (2003). "Experimental determination and system level analysis of essential genes in Escherichia coli MG1655." J Bacteriol 185(19);5673-84. PMID: 13129938

GOA01: 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

Hunt94: Hunt MD, Pettis GS, McIntosh MA (1994). "Promoter and operator determinants for fur-mediated iron regulation in the bidirectional fepA-fes control region of the Escherichia coli enterobactin gene system." J Bacteriol 176(13);3944-55. PMID: 8021177

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

Joyce06: Joyce AR, Reed JL, White A, Edwards R, Osterman A, Baba T, Mori H, Lesely SA, Palsson BO, Agarwalla S (2006). "Experimental and computational assessment of conditionally essential genes in Escherichia coli." J Bacteriol 188(23);8259-71. PMID: 17012394

Lambalot96: Lambalot RH, Gehring AM, Flugel RS, Zuber P, LaCelle M, Marahiel MA, Reid R, Khosla C, Walsh CT (1996). "A new enzyme superfamily - the phosphopantetheinyl transferases." Chem Biol 1996;3(11);923-36. PMID: 8939709

Pettis87: Pettis GS, McIntosh MA (1987). "Molecular characterization of the Escherichia coli enterobactin cistron entF and coupled expression of entF and the fes gene." J Bacteriol 1987;169(9);4154-62. PMID: 3040679

Reichert92: Reichert J, Sakaitani M, Walsh CT (1992). "Characterization of EntF as a serine-activating enzyme." Protein Sci 1992;1(4);549-56. PMID: 1338974

Rusnak91: Rusnak F, Sakaitani M, Drueckhammer D, Reichert J, Walsh CT (1991). "Biosynthesis of the Escherichia coli siderophore enterobactin: sequence of the entF gene, expression and purification of EntF, and analysis of covalent phosphopantetheine." Biochemistry 1991;30(11);2916-27. PMID: 1826089

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

UniProt10: UniProt Consortium (2010). "UniProt version 2010-11 released on 2010-11-02 00:00:00." Database.

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

UniProt11: UniProt Consortium (2011). "UniProt version 2011-06 released on 2011-06-30 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

Other References Related to Gene Regulation

Chen07a: Chen Z, Lewis KA, Shultzaberger RK, Lyakhov IG, Zheng M, Doan B, Storz G, Schneider TD (2007). "Discovery of Fur binding site clusters in Escherichia coli by information theory models." Nucleic Acids Res 35(20);6762-77. PMID: 17921503

Constantinidou06: Constantinidou C, Hobman JL, Griffiths L, Patel MD, Penn CW, Cole JA, Overton TW (2006). "A reassessment of the FNR regulon and transcriptomic analysis of the effects of nitrate, nitrite, NarXL, and NarQP as Escherichia coli K12 adapts from aerobic to anaerobic growth." J Biol Chem 281(8);4802-15. PMID: 16377617

Escolar98: Escolar L, Perez-Martin J, de Lorenzo V (1998). "Coordinated repression in vitro of the divergent fepA-fes promoters of Escherichia coli by the iron uptake regulation (Fur) protein." J Bacteriol 180(9);2579-82. PMID: 9573216

Kumar11: Kumar R, Shimizu K (2011). "Transcriptional regulation of main metabolic pathways of cyoA, cydB, fnr, and fur gene knockout Escherichia coli in C-limited and N-limited aerobic continuous cultures." Microb Cell Fact 10;3. PMID: 21272324

Newman99: Newman DL, Shapiro JA (1999). "Differential fiu-lacZ fusion regulation linked to Escherichia coli colony development." Mol Microbiol 33(1);18-32. PMID: 10411720

Pettis88: Pettis GS, Brickman TJ, McIntosh MA (1988). "Transcriptional mapping and nucleotide sequence of the Escherichia coli fepA-fes enterobactin region. Identification of a unique iron-regulated bidirectional promoter." J Biol Chem 263(35);18857-63. PMID: 2974033

Vassinova00: Vassinova N, Kozyrev D (2000). "A method for direct cloning of fur-regulated genes: identification of seven new fur-regulated loci in Escherichia coli." Microbiology 146 Pt 12;3171-82. PMID: 11101675

WhiteZiegler07: White-Ziegler CA, Malhowski AJ, Young S (2007). "Human body temperature (37degrees C) increases the expression of iron, carbohydrate, and amino acid utilization genes in Escherichia coli K-12." J Bacteriol 189(15);5429-40. PMID: 17526711

Zhang05a: Zhang Z, Gosset G, Barabote R, Gonzalez CS, Cuevas WA, Saier MH (2005). "Functional interactions between the carbon and iron utilization regulators, Crp and Fur, in Escherichia coli." J Bacteriol 187(3);980-90. PMID: 15659676


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Please cite the following article in publications resulting from the use of EcoCyc: Nucleic Acids Research 41:D605-12 2013
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