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Literature summary for 4.1.99.17 extracted from

  • Coquille, S.; Roux, C.; Mehta, A.; Begley, T.P.; Fitzpatrick, T.B.; Thore, S.
    High-resolution crystal structure of the eukaryotic HMP-P synthase (THIC) from Arabidopsis thaliana (2013), J. Struct. Biol., 184, 438-444.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
gene thiC, recombinant expression of His-tagged N-terminally truncated mutant DELTAN71-AtTHIC in Escherichia coli strain BL21(DE3) Arabidopsis thaliana

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant His-tagged DELTAN71-AtTHIC mutant, by a proteolysis assay (especially alpha-chymotrypsin) coupled with the sitting drop vapor diffusion technique at 18°C, using 0.01 M cobalt (II) chloride hexahydrate, 0.1 M sodium acetate trihydrate, pH 4.6, 1 M 1,6-hexanediol, X-ray diffraction structure determination and analysis at 1.6 A resolution Arabidopsis thaliana

Protein Variants

Protein Variants Comment Organism
additional information generation of an N-terminally truncated version of Arabidopsis thaliana THIC, lacking the first 71 amino acids, a chloroplastidial targeting peptide. The truncated AtTHIC is functionally active Arabidopsis thaliana

Localization

Localization Comment Organism GeneOntology No. Textmining
chloroplast
-
Arabidopsis thaliana 9507
-

Metals/Ions

Metals/Ions Comment Organism Structure
Co2+ a metal ion with octahedral coordination (two strictly conserved histidine residues (H426 and H490) and four water molecules) at the same location as a zinc ion in the bacterial enzyme, and a metal ion with multiple coordinated water molecules in the close vicinity of the substrate binding sites, binding structures, overview Arabidopsis thaliana
Fe2+ the enzyme contains a [4Fe-4S] cluster Arabidopsis thaliana

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
65451
-
2 * 65451, recombinant His-tagged DELTAN71-AtTHIC mutant, mass spectrometry and crystal structure Arabidopsis thaliana
130000
-
recombinant His-tagged DELTAN71-AtTHIC mutant, gel filtration Arabidopsis thaliana

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
5-amino-1-(5-phospho-D-ribosyl)imidazole + S-adenosyl-L-methionine Arabidopsis thaliana
-
4-amino-2-methyl-5-phosphomethylpyrimidine + 5'-deoxyadenosine + L-methionine + formate + CO
-
?

Organism

Organism UniProt Comment Textmining
Arabidopsis thaliana O82392 gene thiC
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged N-terminally truncated mutant DELTAN71-AtTHIC from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Arabidopsis thaliana

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5-amino-1-(5-phospho-D-ribosyl)imidazole + S-adenosyl-L-methionine
-
Arabidopsis thaliana 4-amino-2-methyl-5-phosphomethylpyrimidine + 5'-deoxyadenosine + L-methionine + formate + CO
-
?
5-amino-1-(5-phospho-D-ribosyl)imidazole + S-adenosyl-L-methionine the enzyme uses an iron-sulfur cluster as well as a 5'-deoxyadenosyl radical as cofactors to rearrange the 5-amino-imidazole ribonucleotide (AIR) substrate to the pyrimidine ring Arabidopsis thaliana 4-amino-2-methyl-5-phosphomethylpyrimidine + 5'-deoxyadenosine + L-methionine + formate + CO
-
?

Subunits

Subunits Comment Organism
dimer 2 * 65451, recombinant His-tagged DELTAN71-AtTHIC mutant, mass spectrometry and crystal structure Arabidopsis thaliana
More enzyme three-dimensional structure analysis, overview Arabidopsis thaliana

Synonyms

Synonyms Comment Organism
HMP-P synthase
-
Arabidopsis thaliana
thiC
-
Arabidopsis thaliana

Cofactor

Cofactor Comment Organism Structure
S-adenosyl-L-methionine a S-adenosyl-L-methionine radical enzyme Arabidopsis thaliana

General Information

General Information Comment Organism
metabolism the enzyme is important in thiamine diphosphate, vitamin B1, biosynthesis, an essential cofactor for key cellular metabolic enzymes in all forms of life Arabidopsis thaliana
additional information enzyme three-dimensional structure analysis and comparison, overview Arabidopsis thaliana