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Escherichia coli K-12 substr. MG1655 Protein: thiazole synthase

Subunit composition of thiazole synthase = [(ThiG)(ThiH)]6
         thiazole synthase = (ThiG)(ThiH)
                 tyrosine lyase = ThiH (extended summary available)

Summary:
The ThiGH complex catalyzes the conversion of 1-deoxy-D-xylulose 5-phosphate to a thiazole as part of thiamine synthesis.

ThiG and ThiH combine to form a complex containing an iron-sulfuer cluster [Leonardi03]. Together, they are required for the synthesis of 4-methyl-5-(β-hydroxyethyl)thiazole phosphate, which is the rate-limiting step in thiamine synthesis [Vander93, Leonardi04].

Molecular Weight: 440.0 kD (experimental) [Challand10 ]

Gene-Reaction Schematic: ?

Credits:
Created 15-Mar-2010 by Caspi R , SRI International


Subunit of thiazole synthase: thiazole synthase

Molecular Weight: 440 kD (experimental) [Leonardi03]

Summary:
The ThiGH complex catalyzes the conversion of 1-deoxy-D-xylulose 5-phosphate to a thiazole as part of thiamine synthesis.

ThiG and ThiH combine to form a complex containing an iron-sulfuer cluster [Leonardi03]. Together, they are required for the synthesis of 4-methyl-5-(β-hydroxyethyl)thiazole phosphate, which is the rate-limiting step in thiamine synthesis [Vander93, Leonardi04].


Component enzyme of thiazole synthase : ThiG

Synonyms: ThiB, ThiG

Gene: thiG Accession Numbers: EG11589 (EcoCyc), b3991, ECK3982

Locations: cytosol

Sequence Length: 256 AAs

Molecular Weight: 26.896 kD (from nucleotide sequence)

Molecular Weight: 26.9 kD (experimental) [Kelleher98]

pI: 8.59

GO Terms:

Biological Process: GO:0009228 - thiamine biosynthetic process Inferred from experiment Inferred by computational analysis [UniProtGOA11a, GOA06, GOA01a, Vander93]
GO:0009229 - thiamine diphosphate biosynthetic process Inferred by computational analysis [UniProtGOA12]
Molecular Function: GO:0005515 - protein binding Inferred from experiment [Leonardi03]
GO:0003824 - catalytic activity Inferred by computational analysis [GOA01a]
GO:0016740 - transferase activity Inferred by computational analysis [UniProtGOA11a]
GO:0016783 - sulfurtransferase activity Inferred by computational analysis [GOA06]
GO:0036355 - 2-iminoacetate synthase activity Inferred by computational analysis [GOA01a]
Cellular Component: GO:0005829 - cytosol Inferred from experiment Inferred by computational analysis [DiazMejia09, Ishihama08, LopezCampistrou05]
GO:0005737 - cytoplasm Inferred by computational analysis [UniProtGOA11, UniProtGOA11a, GOA06]

MultiFun Terms: metabolism biosynthesis of building blocks cofactors, small molecule carriers thiamin

Unification Links: DIP:DIP-6868N , EcoliWiki:b3991 , Mint:MINT-1289283 , ModBase:P30139 , PR:PRO_000024056 , Protein Model Portal:P30139 , RefSeq:NP_418418 , SMR:P30139 , String:511145.b3991 , Swiss-Model:P30139 , UniProt:P30139

Relationship Links: InterPro:IN-FAMILY:IPR008867 , InterPro:IN-FAMILY:IPR013785 , Pfam:IN-FAMILY:PF05690

Catalyzes:
1-deoxy-D-xylulose 5-phosphate + 2-iminoacetate + a thiocarboxy-adenylated-[ThiS-Protein] → 2-[(2R,5Z)-(2-carboxy-4-methylthiazol-5(2H)-ylidene]ethyl phosphate + a ThiS sulfur-carrier protein + 2 H2O

Summary:
ThiG is involved in the synthesis of the thiazole moiety of thiamin. Thiazole synthase is a heterodimer of ThiG and ThiH, which catalyzes the conversion of 1-deoxy-D-xylulose 5-phosphate to a thiazole. [Leonardi03] ThiG and ThiH complex is required for the synthesis of 4-methyl-5-(beta-hydroxylethyl) thiazole phosphate, which is the rate-limiting step in thiamine synthesis. The formation of 4-methyl-5-(-hydroxyethyl)thiazole phosphate requires the precursor 1-deoxyxyulose 5-phosphate, a sulfur atom from cysteine which is transferred via ThiS, and dehydroglycine. [Vander93, Leonardi04, Dorrestein04]

ThiG and ThiH copurify as a large multimeric non-covalent complex. ThiG catalyzes the thiazole cyclization reaction and ThiH is a tyrosine lyase that cleaves tyrosine to form dehydroglycine. The activity of thiazole phosphate forming reaction using purified protein components required four proteins isolated as heterodimers, ThiGH and ThiFS. Experimental evidence indicated a presence of an iron-sulfur cluster within this complex and that the S-adenosylmethionine (AdoMet) bound this cluster. [Kriek07, Leonardi03]

Essentiality data for thiG knockouts: ?

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

Component enzyme of thiazole synthase : ThiH

Synonyms: ThiB, ThiH

Gene: thiH Accession Numbers: EG11590 (EcoCyc), b3990, ECK3981

Locations: cytosol

Sequence Length: 377 AAs

Molecular Weight: 43.32 kD (from nucleotide sequence)

Molecular Weight: 42.0 kD (experimental) [Challand10]

pI: 7.0

GO Terms:

Biological Process: GO:0006974 - cellular response to DNA damage stimulus Inferred from experiment [Khil02]
GO:0009228 - thiamine biosynthetic process Inferred from experiment Inferred by computational analysis [UniProtGOA11a, GOA01a, Vander93]
GO:0009229 - thiamine diphosphate biosynthetic process Inferred by computational analysis [UniProtGOA12]
Molecular Function: GO:0005515 - protein binding Inferred from experiment [Leonardi03]
GO:0016829 - lyase activity Inferred from experiment Inferred by computational analysis [UniProtGOA11a, Challand10]
GO:0051536 - iron-sulfur cluster binding Inferred from experiment Inferred by computational analysis [UniProtGOA11a, GOA01a, Kriek07]
GO:0051539 - 4 iron, 4 sulfur cluster binding Inferred from experiment Inferred by computational analysis [UniProtGOA11a, GOA01a, Kriek07]
GO:0003824 - catalytic activity Inferred by computational analysis [GOA01a]
GO:0005506 - iron ion binding Inferred by computational analysis [GOA01a]
GO:0036355 - 2-iminoacetate synthase activity Inferred by computational analysis [GOA01]
GO:0046872 - metal ion binding Inferred by computational analysis [UniProtGOA11a]
Cellular Component: GO:0005829 - cytosol Inferred by computational analysis [DiazMejia09]

MultiFun Terms: metabolism biosynthesis of building blocks cofactors, small molecule carriers thiamin

Unification Links: DIP:DIP-6869N , EcoliWiki:b3990 , ModBase:P30140 , PR:PRO_000024057 , Pride:P30140 , Protein Model Portal:P30140 , RefSeq:NP_418417 , SMR:P30140 , String:511145.b3990 , UniProt:P30140

Relationship Links: InterPro:IN-FAMILY:IPR007197 , InterPro:IN-FAMILY:IPR010722 , InterPro:IN-FAMILY:IPR012726 , InterPro:IN-FAMILY:IPR013785 , Pfam:IN-FAMILY:PF04055 , Pfam:IN-FAMILY:PF06968 , Smart:IN-FAMILY:SM00876

Catalyzes:
L-tyrosine + S-adenosyl-L-methionine + NADPH → 2-iminoacetate + 4-methylphenol + 5'-deoxyadenosine + L-methionine + NADP+ + H+

Summary:
ThiH is a tyrosine lyase that cleaves the Cα-Cβ bond of L-tyrosine, generating 4-methylphenol as a by-product, to form 2-iminoacetate. The later acts as one of three substrates for the thiazole cyclization reaction catalyzed by ThiG. [Challand10]

ThiH is a radical S-adenosylmethionine enzyme that utilizes a [4Fe-4S] cluster to reductively cleave AdoMet, forming methionine and a 5'-deoxyadenosyl radical. The radical abstracts a hydrogen radical from a residue in ThiH. Once radicalyzed, the enzyme attach L-tyrosine, forming 2-iminoacetate and 4-methylphenol [Kriek07a, Kriek07].

ThiH and ThiG form a complex. It is possible that 2-iminoacetate remains bound to the enzyme, ensuring that tyrosine cleavage catalyzed by ThiH is synchronized with the incorporation of dehydroglycine by ThiG into the thiazole carboxylate [Kriek07a].

Mutagenesis of the thiH gene was performed and residues required for in vivo function were identified. [MartinezGomez04]

Essentiality data for thiH knockouts: ?

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

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

Challand10: Challand MR, Martins FT, Roach PL (2010). "Catalytic activity of the anaerobic tyrosine lyase required for thiamine biosynthesis in Escherichia coli." J Biol Chem 285(8);5240-8. PMID: 19923213

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

Dorrestein04: Dorrestein PC, Zhai H, McLafferty FW, Begley TP (2004). "The biosynthesis of the thiazole phosphate moiety of thiamin: the sulfur transfer mediated by the sulfur carrier protein ThiS." Chem Biol 11(10);1373-81. PMID: 15489164

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

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."

GOA06: GOA, SIB (2006). "Electronic Gene Ontology annotations created by transferring manual GO annotations between orthologous microbial proteins."

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

Kelleher98: Kelleher NL, Taylor SV, Grannis D, Kinsland C, Chiu HJ, Begley TP, McLafferty FW (1998). "Efficient sequence analysis of the six gene products (7-74 kDa) from the Escherichia coli thiamin biosynthetic operon by tandem high-resolution mass spectrometry." Protein Sci 7(8);1796-801. PMID: 10082377

Khil02: Khil PP, Camerini-Otero RD (2002). "Over 1000 genes are involved in the DNA damage response of Escherichia coli." Mol Microbiol 44(1);89-105. PMID: 11967071

Kriek07: Kriek M, Martins F, Leonardi R, Fairhurst SA, Lowe DJ, Roach PL (2007). "Thiazole synthase from Escherichia coli: an investigation of the substrates and purified proteins required for activity in vitro." J Biol Chem 282(24);17413-23. PMID: 17403671

Kriek07a: Kriek M, Martins F, Challand MR, Croft A, Roach PL (2007). "Thiamine biosynthesis in Escherichia coli: identification of the intermediate and by-product derived from tyrosine." Angew Chem Int Ed Engl 46(48);9223-6. PMID: 17969213

Leonardi03: Leonardi R, Fairhurst SA, Kriek M, Lowe DJ, Roach PL (2003). "Thiamine biosynthesis in Escherichia coli: isolation and initial characterisation of the ThiGH complex." FEBS Lett 539(1-3);95-9. PMID: 12650933

Leonardi04: Leonardi R, Roach PL (2004). "Thiamine biosynthesis in Escherichia coli: in vitro reconstitution of the thiazole synthase activity." J Biol Chem 279(17);17054-62. PMID: 14757766

LopezCampistrou05: Lopez-Campistrous A, Semchuk P, Burke L, Palmer-Stone T, Brokx SJ, Broderick G, Bottorff D, Bolch S, Weiner JH, Ellison MJ (2005). "Localization, annotation, and comparison of the Escherichia coli K-12 proteome under two states of growth." Mol Cell Proteomics 4(8);1205-9. PMID: 15911532

MartinezGomez04: Martinez-Gomez NC, Robers M, Downs DM (2004). "Mutational analysis of ThiH, a member of the radical S-adenosylmethionine (AdoMet) protein superfamily." J Biol Chem 279(39);40505-10. PMID: 15271986

UniProtGOA11: UniProt-GOA (2011). "Gene Ontology annotation based on the manual assignment of UniProtKB Subcellular Location terms in UniProtKB/Swiss-Prot entries."

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."

Vander93: Vander Horn PB, Backstrom AD, Stewart V, Begley TP (1993). "Structural genes for thiamine biosynthetic enzymes (thiCEFGH) in Escherichia coli K-12." J Bacteriol 1993;175(4);982-92. PMID: 8432721


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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 Dec 19, 2014, biocyc14.