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Escherichia coli K-12 substr. MG1655 Polypeptide: IlvI



Gene: ilvI Accession Numbers: EG10500 (EcoCyc), b0077, ECK0079

Regulation Summary Diagram: ?

Component of: acetolactate synthase / acetohydroxybutanoate synthase (extended summary available)

Summary:
IlvI is the large catalytic subunit of the bifunctional acetohydroxybutanoate synthase /acetolactate synthase (IlvI/H, AHAS III) which carries out both the first step in valine biosynthesis and the second step in isoleucine biosynthesis. The IlvI/H protein complex catalyzes the conversion of pyruvate and oxobutanoate into 2-aceto-2-hydroxy-butyrate and the conversion of pyruvate into 2-acetolactate. Both reactions generate carbon dioxide as a product. [De74, Gollop89, Barak87, Vyazmensky96, Weinstock92]

C-terminal and N-terminal ilvH mutants were constructed and used to determine the minimum activation peptide necessary to activate IlvI [Zhao13]. Interactions between large and small subunits of different AHAS isozymes were investigated. IlvI could be activated by IlvM just as well as IlvH. [Vyazmensky09]

Isolated IlvI has only 5% of the molar activity of its holoenzyme. However, isolated IlvI has similar substrate specificity and similar cofactor dependence as its holoenzyme. Assembly of the holoenzyme requires FAD. [Vyazmensky96]

Gene Citations: [Haughn85, Blattner97, Jafri02, Wang93]

Locations: cytosol

Map Position: [85,630 -> 87,354] (1.85 centisomes)
Length: 1725 bp / 574 aa

Molecular Weight of Polypeptide: 62.984 kD (from nucleotide sequence), 60 kD (experimental) [Vyazmensky96 ]

Isozyme Sequence Similarity [Comment 1]:
acetolactate synthase II, large subunit, C-ter fragment (pseudogene): YES ,
acetohydroxy acid synthase I, large subunit: YES

Unification Links: ASAP:ABE-0000290 , CGSC:601 , DIP:DIP-6850N , EchoBASE:EB0495 , EcoGene:EG10500 , EcoliWiki:b0077 , ModBase:P00893 , OU-Microarray:b0077 , PortEco:ilvI , PR:PRO_000023008 , Pride:P00893 , Protein Model Portal:P00893 , RefSeq:YP_025294 , RegulonDB:EG10500 , SMR:P00893 , String:511145.b0077 , UniProt:P00893

Relationship Links: InterPro:IN-FAMILY:IPR000399 , InterPro:IN-FAMILY:IPR011766 , InterPro:IN-FAMILY:IPR012000 , InterPro:IN-FAMILY:IPR012001 , InterPro:IN-FAMILY:IPR012846 , Panther:IN-FAMILY:PTHR18968:SF13 , Pfam:IN-FAMILY:PF00205 , Pfam:IN-FAMILY:PF02775 , Pfam:IN-FAMILY:PF02776 , Prosite:IN-FAMILY:PS00187

In Paralogous Gene Group: 144 (4 members)

Gene-Reaction Schematic: ?

Genetic Regulation Schematic: ?

GO Terms:

Biological Process: GO:0009097 - isoleucine biosynthetic process Inferred from experiment Inferred by computational analysis [UniProtGOA12, Gollop89]
GO:0009099 - valine biosynthetic process Inferred from experiment Inferred by computational analysis [UniProtGOA12, Gollop89]
GO:0008652 - cellular amino acid biosynthetic process Inferred by computational analysis [UniProtGOA11a]
GO:0009082 - branched-chain amino acid biosynthetic process Inferred by computational analysis [UniProtGOA11a, GOA01a]
Molecular Function: GO:0000287 - magnesium ion binding Inferred from experiment Inferred by computational analysis [GOA01a, Vyazmensky96]
GO:0003984 - acetolactate synthase activity Inferred from experiment Inferred by computational analysis [GOA01, GOA01a, De74, Gollop89]
GO:0030976 - thiamine pyrophosphate binding Inferred from experiment Inferred by computational analysis [GOA01a, Vyazmensky96]
GO:0050660 - flavin adenine dinucleotide binding Inferred from experiment Inferred by computational analysis [GOA01a, Vyazmensky96]
GO:0003824 - catalytic activity Inferred by computational analysis [GOA01a]
GO:0016740 - transferase activity Inferred by computational analysis [UniProtGOA11a]
GO:0046872 - metal ion binding Inferred by computational analysis [UniProtGOA11a]
Cellular Component: GO:0005829 - cytosol Inferred from experiment Inferred by computational analysis [DiazMejia09, Ishihama08]

MultiFun Terms: metabolism biosynthesis of building blocks amino acids isoleucine/valine

Essentiality data for ilvI knockouts: ?

Growth Medium Growth? T (°C) O2 pH Osm/L Growth Observations
LB enriched Yes 37 Aerobic 6.95   Yes [Gerdes03, Comment 2]
LB Lennox Yes 37 Aerobic 7   Yes [Baba06, Comment 3]
M9 medium with 1% glycerol Yes 37 Aerobic 7.2 0.35 Yes [Joyce06, Comment 4]
MOPS medium with 0.4% glucose Yes 37 Aerobic 7.2 0.22 Yes [Baba06, Comment 3]
Yes [Feist07, Comment 5]

Credits:
Curated 08-Feb-2006 by Keseler I , SRI International
Last-Curated ? 14-Oct-2013 by Kubo A , SRI International


Subunit of: acetolactate synthase / acetohydroxybutanoate synthase

Synonyms: acetolactate synthase III, acetohydroxy acid synthase III, AHAS III, acetohydroxybutanoate synthase III

Subunit composition of acetolactate synthase / acetohydroxybutanoate synthase = [IlvI]2[IlvH]2

Summary:
Acetohydroxy acid synthase III (AHAS III) is one of two functional isozymes catalyzing the decarboxylation of pyruvate and transfer of the resulting acetaldehyde group to either pyruvate or α-ketobutyrate, producing α-acetolactate for the valine pathway and α-aceto-α-hydroxybutyrate for the isoleucine pathway. This is the first common step in the biosynthesis of the branched-chain amino acids isoleucine, leucine, and valine. A third isozyme, AHAS II, is not functional in E. coli K-12 due to the presence of a frame shift mutation in the gene encoding the large subunit, ilvG. [Neidhardt96]

In the presence of both pyruvate and α-ketobutyrate, AHAS III produces approximately 40-fold more acetohydroxybutyrate than acetolactate, while AHAS I shows no product preference [Barak87, Gollop89].

The differential regulation of enzymatic activity and expression of the isozymes has direct physiological consequences and has been under intense study. The end products of the branched-chain amino acid biosynthesis pathways all inhibit AHAS III activity, although inhibition by valine is most significant [Gollop89]. Both AHAS I and III are inhibited by valine [De78]. Activity of AHAS III is only partially inhibited by leucine, while AHAS I activity can be almost completely inhibited [Gollop83].

Molecular Weight: 155 kD (experimental) [Vyazmensky96]

Credits:
Last-Curated ? 29-Aug-2007 by Shearer A , SRI International


Enzymatic reaction of: acetolactate synthase

Synonyms: acetolactate pyruvate-lyase (carboxylating), acetohydroxy acid synthase

EC Number: 2.2.1.6

2 pyruvate + H+ <=> (S)-2-acetolactate + CO2

The reaction direction shown, that is, A + B ↔ C + D versus C + D ↔ A + B, is in accordance with the Enzyme Commission system.

This reaction is reversible.

In Pathways: superpathway of leucine, valine, and isoleucine biosynthesis , valine biosynthesis

Cofactors or Prosthetic Groups: thiamin diphosphate [De78, Vyazmensky96], FAD [Vyazmensky96], Mg2+ [Vyazmensky96]

Inhibitors (Unknown Mechanism): sulfometuron methyl [Barak88] , L-isoleucine [Barak88] , L-valine [Gollop89] , L-leucine

Primary Physiological Regulators of Enzyme Activity: L-isoleucine , L-valine , L-leucine

Kinetic Parameters:

Substrate
Km (μM)
Citations
pyruvate
6000.0
[Gollop89]

pH(opt): 9 [De78]


Enzymatic reaction of: acetohydroxybutanoate synthase

Synonyms: acetohydroxybutyrate pyruvate-lyase (carboxylating)

EC Number: 2.2.1.6

pyruvate + 2-oxobutanoate + H+ <=> (S)-2-aceto-2-hydroxybutanoate + CO2

The reaction direction shown, that is, A + B ↔ C + D versus C + D ↔ A + B, is in accordance with the Enzyme Commission system.

The reaction is favored in the direction shown.

In Pathways: superpathway of leucine, valine, and isoleucine biosynthesis , superpathway of threonine metabolism , isoleucine biosynthesis I (from threonine)

Cofactors or Prosthetic Groups: thiamin diphosphate [Vyazmensky96], Mg2+ [Vyazmensky96], FAD [Vyazmensky96]

Inhibitors (Unknown Mechanism): L-valine [Gollop89, Comment 6] , L-leucine [Gollop83]

Kinetic Parameters:

Substrate
Km (μM)
Citations
pyruvate
7600.0
[De78]


Sequence Features

Feature Class Location Citations Comment
Amino-Acid-Sites-That-Bind 51
[UniProt10a]
UniProt: Thiamine pyrophosphate; Non-Experimental Qualifier: by similarity;
Amino-Acid-Sites-That-Bind 153
[UniProt10a]
UniProt: FAD; Non-Experimental Qualifier: by similarity;
Sequence-Conflict 202 -> 203
[Ayala91, Squires83, UniProt10]
Alternate sequence: TL → SV; UniProt: (in Ref. 1; CAA25755 and 5; CAA38854);
Sequence-Conflict 206
[Ayala91, Squires83, UniProt10]
Alternate sequence: A → V; UniProt: (in Ref. 1; CAA25755 and 5; CAA38854);
Sequence-Conflict 254
[Ayala91, Squires83, UniProt10]
Alternate sequence: A → V; UniProt: (in Ref. 1; CAA25755 and 5; CAA38854);
Nucleotide-Phosphate-Binding-Region 261 -> 282
[UniProt10a]
UniProt: FAD; Non-Experimental Qualifier: by similarity;
Nucleotide-Phosphate-Binding-Region 304 -> 323
[UniProt10a]
UniProt: FAD; Non-Experimental Qualifier: by similarity;
Protein-Segment 397 -> 477
[UniProt10a]
UniProt: Thiamine pyrophosphate binding; Sequence Annotation Type: region of interest;
Sequence-Conflict 422
[Ayala91, Squires83, UniProt10]
Alternate sequence: T → S; UniProt: (in Ref. 1; CAA25755 and 5; CAA38854);
Sequence-Conflict 437 -> 438
[Ayala91, UniProt10]
Alternate sequence: LP → FA; UniProt: (in Ref. 5; CAA38854);
Metal-Binding-Site 448
[UniProt10a]
UniProt: Magnesium; Non-Experimental Qualifier: by similarity;
Metal-Binding-Site 475
[UniProt10a]
UniProt: Magnesium; Non-Experimental Qualifier: by similarity;
Sequence-Conflict 507 -> 509
[Ayala91, Squires83, UniProt10]
Alternate sequence: LAE → RG; UniProt: (in Ref. 1; CAA25755 and 5; CAA38854);


Gene Local Context (not to scale): ?

Transcription Units:

Notes:

History:
Socorro Gama-Castro on Fri Oct 29, 2004:
The start site of this gene was originally assigned solely on the basis of sequence considerations [Blattner97 ]. However, it was changed because Lago et al [Lago85 ] attested that the real start site is actually located 90 bp downstream. The demonstration is based on protein sequencing.
10/20/97 Gene b0077 from Blattner lab Genbank (v. M52) entry merged into EcoCyc gene EG10500; confirmed by SwissProt match.


References

AlexanderCaudle90: Alexander-Caudle C, Latinwo LM, Jackson JH (1990). "Acetohydroxy acid synthase activity from a mutation at ilvF in Escherichia coli K-12." J Bacteriol 1990;172(6);3060-5. PMID: 2188950

Ayala91: Ayala J.A. (1991). "Regulation of transcription at the 2-minute region of the genetic map of Escherichia coli." Data submission to EMBL/GenBank/DDBJ databases on 1991-01.

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

Bairoch93: Bairoch A, Boeckmann B (1993). "The SWISS-PROT protein sequence data bank, recent developments." Nucleic Acids Res. 21:3093-3096. PMID: 8332529

Barak87: Barak Z, Chipman DM, Gollop N (1987). "Physiological implications of the specificity of acetohydroxy acid synthase isozymes of enteric bacteria." J Bacteriol 169(8);3750-6. PMID: 3301814

Barak88: Barak Z, Calvo JM, Schloss JV (1988). "Acetolactate synthase isozyme III from Escherichia coli." Methods Enzymol 1988;166;455-8. PMID: 3071721

Blattner97: Blattner FR, Plunkett G, Bloch CA, Perna NT, Burland V, Riley M, Collado-Vides J, Glasner JD, Rode CK, Mayhew GF, Gregor J, Davis NW, Kirkpatrick HA, Goeden MA, Rose DJ, Mau B, Shao Y (1997). "The complete genome sequence of Escherichia coli K-12." Science 277(5331);1453-74. PMID: 9278503

De74: De Felice M, Guardiola J, Esposito B, Iaccarino M (1974). "Structural genes for a newly recognized acetolactate synthase in Escherichia coli K-12." J Bacteriol 120(3);1068-77. PMID: 4612003

De78: De Felice M, Squires C, Levinthal M (1978). "A comparative study of the acetohydroxy acid synthase isoenzymes of Escherichia coli K-12." Biochim. Biophys. Acta 541;9-17.

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

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

Friden85: Friden P, Donegan J, Mullen J, Tsui P, Freundlich M, Eoyang L, Weber R, Silverman PM (1985). "The ilvB locus of Escherichia coli K-12 is an operon encoding both subunits of acetohydroxyacid synthase I." Nucleic Acids Res 1985;13(11);3979-93. PMID: 2989781

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, 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."

Gollop83: Gollop N, Chipman DM, Barak Z (1983). "Inhibition of acetohydroxy acid synthase by leucine." Biochim Biophys Acta 1983;748(1);34-9. PMID: 6351926

Gollop89: Gollop N, Damri B, Barak Z, Chipman DM (1989). "Kinetics and mechanism of acetohydroxy acid synthase isozyme III from Escherichia coli." Biochemistry 28(15);6310-7. PMID: 2675968

Haughn85: Haughn GW, Squires CH, De Felice M, Largo CT, Calvo JM (1985). "Unusual organization of the ilvIH promoter of Escherichia coli." J Bacteriol 1985;163(1);186-98. PMID: 3891724

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

Jafri02: Jafri S, Chen S, Calvo JM (2002). "ilvIH operon expression in Escherichia coli requires Lrp binding to two distinct regions of DNA." J Bacteriol 184(19);5293-300. PMID: 12218014

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

Lago85: Lago CT, Sannia G, Marino G, Squires CH, Calvo JM, De Felice M (1985). "The ilvIH operon of Escherichia coli K-12. Identification of the gene products and recognition of the translational start by polypeptide microsequencing." Biochim Biophys Acta 824(1);74-9. PMID: 3881131

Lawther87: Lawther RP, Wek RC, Lopes JM, Pereira R, Taillon BE, Hatfield GW (1987). "The complete nucleotide sequence of the ilvGMEDA operon of Escherichia coli K-12." Nucleic Acids Res 1987;15(5);2137-55. PMID: 3550695

Neidhardt96: Neidhardt FC, Curtiss III R, Ingraham JL, Lin ECC, Low Jr KB, Magasanik B, Reznikoff WS, Riley M, Schaechter M, Umbarger HE "Escherichia coli and Salmonella, Cellular and Molecular Biology, Second Edition." American Society for Microbiology, Washington, D.C., 1996.

Robinson82: Robinson CL, Jackson JH (1982). "New acetohydroxy acid synthase activity from mutational activation of a cryptic gene in Escherichia coli K-12." Mol Gen Genet 1982;186(2);240-6. PMID: 7050630

Squires83: Squires CH, De Felice M, Devereux J, Calvo JM (1983). "Molecular structure of ilvIH and its evolutionary relationship to ilvG in Escherichia coli K12." Nucleic Acids Res 1983;11(15);5299-313. PMID: 6308579

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.

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."

Vyazmensky09: Vyazmensky M, Zherdev Y, Slutzker A, Belenky I, Kryukov O, Barak Z, Chipman DM (2009). "Interactions between large and small subunits of different acetohydroxyacid synthase isozymes of Escherichia coli." Biochemistry 48(36);8731-7. PMID: 19653643

Vyazmensky96: Vyazmensky M, Sella C, Barak Z, Chipman DM (1996). "Isolation and characterization of subunits of acetohydroxy acid synthase isozyme III and reconstitution of the holoenzyme." Biochemistry 35(32);10339-46. PMID: 8756689

Wang93: Wang Q, Calvo JM (1993). "Lrp, a global regulatory protein of Escherichia coli, binds co-operatively to multiple sites and activates transcription of ilvIH." J Mol Biol 1993;229(2);306-18. PMID: 8429549

Weinstock92: Weinstock O, Sella C, Chipman DM, Barak Z (1992). "Properties of subcloned subunits of bacterial acetohydroxy acid synthases." J Bacteriol 1992;174(17);5560-6. PMID: 1512191

Zhao13: Zhao Y, Niu C, Wen X, Xi Z (2013). "The minimum activation peptide from ilvH can activate the catalytic subunit of AHAS from different species." Chembiochem 14(6);746-52. PMID: 23512804

Other References Related to Gene Regulation

Levinthal94: Levinthal M, Lejeune P, Danchin A (1994). "The H-NS protein modulates the activation of the ilvIH operon of Escherichia coli K12 by Lrp, the leucine regulatory protein." Mol Gen Genet 242(6);736-43. PMID: 8152423

Marasco94: Marasco R, Varcamonti M, La Cara F, Ricca E, De Felice M, Sacco M (1994). "In vivo footprinting analysis of Lrp binding to the ilvIH promoter region of Escherichia coli." J Bacteriol 176(17);5197-201. PMID: 8071194

Platko90: Platko JV, Willins DA, Calvo JM (1990). "The ilvIH operon of Escherichia coli is positively regulated." J Bacteriol 172(8);4563-70. PMID: 2115869

Platko93: Platko JV, Calvo JM (1993). "Mutations affecting the ability of Escherichia coli Lrp to bind DNA, activate transcription, or respond to leucine." J Bacteriol 175(4);1110-7. PMID: 8432705

Ricca89: Ricca E, Aker DA, Calvo JM (1989). "A protein that binds to the regulatory region of the Escherichia coli ilvIH operon." J Bacteriol 171(3);1658-64. PMID: 2646291

Wang93a: Wang Q, Calvo JM (1993). "Lrp, a major regulatory protein in Escherichia coli, bends DNA and can organize the assembly of a higher-order nucleoprotein structure." EMBO J 12(6);2495-501. PMID: 8508774

Wang93b: Wang Q, Sacco M, Ricca E, Lago CT, De Felice M, Calvo JM (1993). "Organization of Lrp-binding sites upstream of ilvIH in Salmonella typhimurium." Mol Microbiol 7(6);883-91. PMID: 8483418

Willins92: Willins DA, Calvo JM (1992). "In vitro transcription from the Escherichia coli ilvIH promoter." J Bacteriol 174(23);7648-55. PMID: 1447135


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Please cite the following article in publications resulting from the use of EcoCyc: Nucleic Acids Research 41:D605-12 2013
Page generated by SRI International Pathway Tools version 18.5 on Fri Nov 28, 2014, biocyc12.