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Information on EC 2.7.7.73 - sulfur carrier protein ThiS adenylyltransferase and Organism(s) Escherichia coli and UniProt Accession P30138

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IUBMB Comments
Binds Zn2+. The enzyme catalyses the adenylation of ThiS, a sulfur carrier protein involved in the biosynthesis of thiamine. The enzyme shows significant structural similarity to ubiquitin-activating enzyme [3,4]. In Escherichia coli, but not in Bacillus subtilis, the enzyme forms a cross link from Cys-184 to the ThiS carboxy terminus (the position that is also thiolated) via an acyldisulfide .
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Escherichia coli
UNIPROT: P30138
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The taxonomic range for the selected organisms is: Escherichia coli
The expected taxonomic range for this enzyme is: Bacteria, Archaea, Eukaryota
Synonyms
ThiF, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
PATHWAY SOURCE
PATHWAYS
-
-, -
SYSTEMATIC NAME
IUBMB Comments
ATP:[ThiS] adenylyltransferase
Binds Zn2+. The enzyme catalyses the adenylation of ThiS, a sulfur carrier protein involved in the biosynthesis of thiamine. The enzyme shows significant structural similarity to ubiquitin-activating enzyme [3,4]. In Escherichia coli, but not in Bacillus subtilis, the enzyme forms a cross link from Cys-184 to the ThiS carboxy terminus (the position that is also thiolated) via an acyldisulfide [2].
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ATP + [sulfur-carrier protein ThiS]
diphosphate + adenylyl-[sulfur-carrier protein ThiS]
show the reaction diagram
ATP + [sulfur-carrier protein ThiS]
diphosphate + adenylyl-[sulfur-carrier protein ThiS]
show the reaction diagram
-
ThiF catalyzes the adenylation of the carboxy terminus of ThiS and the subsequent displacement of AMP catalyzed by ThiI-persulfide to give a ThiS-ThiI acyl disulfide
-
-
?
additional information
?
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ThiF shows no activity with GTP, CTP or TTP as cosubstrates
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-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
ATP + [sulfur-carrier protein ThiS]
diphosphate + adenylyl-[sulfur-carrier protein ThiS]
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Zn2+
each monomer of ThiF binds one zinc atom
Zn2+
-
ThiF contains a Zn-sulfur center
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
ThiF is an enzyme in the biosynthetic cascade for generating the essential cofactor thiamin diphosphate
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
27000
-
2 * 27000, calculated from amino acid sequence
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
x-ray crystallography
homodimer
-
2 * 27000, calculated from amino acid sequence
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
ThiF alone and in complex with ATP, hanging drop vapor diffusion method, using 14% (w/v) ammonium acetate, 50 mM MES (pH 6.5), 5 mM dithiothreitol, at 25°C
in complex with ThiS, hanging drop vapor diffusion method, using 7-8% polyethylene glycol 400, 35 mM calcium chloride, and 100 mM Tris-HCl (pH 7.0-7.3)
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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C184S
inactive, C184S is equally effective in catalyzing the formation of [ThiS]-COSH as the wild type enzyme. Cys184 is directly involved in the crosslinking of ThiF to ThiS but is not essential for the formation of [ThiS]-COSH
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate precipitation, DEAE column chromatography, and Superdex 75 gel filtration
G-75 Sephadex gel filtration
glutathione affinity column chromatography, Resource Q column chromatography, and Superdex 200 gel filtration
Hi-Trap QFF column chromatography and Superdex 200 gel filtration
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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli BL21(DE3) cells
expressed in Escherichia coli BL834(DE3) cells
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REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Lehmann, C.; Begley, T.P.; Ealick, S.E.
Structure of the Escherichia coli ThiS-ThiF complex, a key component of the sulfur transfer system in thiamin biosynthesis
Biochemistry
45
11-19
2006
Escherichia coli
Manually annotated by BRENDA team
Taylor, S.V.; Kelleher, N.L.; Kinsland, C.; Chiu, H.J.; Costello, C.A.; Backstrom, A.D.; McLafferty, F.W.; Begley, T.P.
Thiamin biosynthesis in Escherichia coli. Identification of ThiS thiocarboxylate as the immediate sulfur donor in the thiazole formation
J. Biol. Chem.
273
16555-16560
1998
Escherichia coli (P30138)
Manually annotated by BRENDA team
Duda, D.M.; Walden, H.; Sfondouris, J.; Schulman, B.A.
Structural analysis of Escherichia coli ThiF
J. Mol. Biol.
349
774-786
2005
Escherichia coli (P30138)
Manually annotated by BRENDA team
Xi, J.; Ge, Y.; Kinsland, C.; McLafferty, F.W.; Begley, T.P.
Biosynthesis of the thiazole moiety of thiamin in Escherichia coli: identification of an acyldisulfide-linked protein--protein conjugate that is functionally analogous to the ubiquitin/E1 complex
Proc. Natl. Acad. Sci. USA
98
8513-8518
2001
Escherichia coli (P30138)
Manually annotated by BRENDA team