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Literature summary for 2.1.1.132 extracted from

  • Keller, J.P.; Smith, P.M.; Benach, J.; Christendat, D.; de Titta, G.T.; Hunt, J.F.
    The crystal structure of MT0146/CbiT suggests that the putative precorrin-8w decarboxylase is a methyltransferase (2002), Structure, 10, 1475-1487.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
gene MT0146 or cbiT, fused to gene cbiE, resulting in cobL, DNA and amino acid sequence determination and analysis, promotor analysis, expression in Escherichia coli strain B834 Methanothermobacter thermautotrophicus

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant apo-enzyme CbiT or in complex with S-adenosyl-L-methionine, hanging drop vapour diffusion method, 25°C, for the apo-enzyme: 5-10 mg/ml protein in 10% w/v PEG 8000, 100 mM MgCl2, and 5 mM Tric-HCl, pH 7.5, 0.002 ml equilibrated against 1 ml of well solution containing 16-22% PEG 8000, 0.2 M MgCl2, 10 mM Tris-HCl, pH 7.5, crystals appear after 1 day, for the binary complex: 1-2 mg/ml protein in 0.5 mM S-adenosyl-L-methionine, 0.1-0.2% PEG 8000, 50 mM MgCl2, 2.5 mM Tris-HCl, pH 7.5, 0.004 ml equilibrated against 1 ml of well solution containing 0.2-0.4% PEG 8000, 0.2 M MgCl2, and 5 mM Tris-HCl, pH 7.5, cryoprotection by soaking in 25% PEG 8000, 0.2 M MgCl2, and glycerol up to 25% w/v, freezing in liquid propane, X-ray diffraction structure determination and analysis at 1.9 A resolution, modeling Methanothermobacter thermautotrophicus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
21000
-
4 * 21000, static light scattering and SDS-PAGE Methanothermobacter thermautotrophicus
81000
-
static light scattering Methanothermobacter thermautotrophicus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
additional information Methanothermobacter thermautotrophicus evolutionary origin of the cbiT ?
-
?
S-adenosyl-L-methionine + cobalt-precorrin-6Y + 6 H+ Methanothermobacter thermautotrophicus steps of the cobalamin, i.e. vitamin B12, biosynthesis, pathway overview S-adenosyl-L-homocysteine + cobalt-precorrin-8X + 2 H2O
-
?

Organism

Organism UniProt Comment Textmining
Methanothermobacter thermautotrophicus
-
gene MT0146 or cbiT, fused to gene cbiE, resulting in cobL
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme CbiT from Escherichia coli strain B834 Methanothermobacter thermautotrophicus

Reaction

Reaction Comment Organism Reaction ID
2 S-adenosyl-L-methionine + precorrin-6B = 2 S-adenosyl-L-homocysteine + precorrin-8X + CO2 CbiT: substrate and cofactor binding, active site structure, mechanism Methanothermobacter thermautotrophicus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information evolutionary origin of the cbiT Methanothermobacter thermautotrophicus ?
-
?
additional information S-adenosyl-L-methionine binding structure and mechanism of CbiT Methanothermobacter thermautotrophicus ?
-
?
S-adenosyl-L-methionine + cobalt-precorrin-6Y + 6 H+ steps of the cobalamin, i.e. vitamin B12, biosynthesis, pathway overview Methanothermobacter thermautotrophicus S-adenosyl-L-homocysteine + cobalt-precorrin-8X + 2 H2O
-
?
S-adenosyl-L-methionine + cobalt-precorrin-6Y + 6 H+ methylation and decarboxylation are catalyzed by 2 different enzyme encoded by gene cbiE, which is responsible for the 2 methylations at C5 and C15, and by gene cbiT, which is responsible for the decarboxylation, the genes are fused together, overview Methanothermobacter thermautotrophicus S-adenosyl-L-homocysteine + cobalt-precorrin-8X + 2 H2O
-
?

Subunits

Subunits Comment Organism
More CbiT has a Rossmann-like methyltransferase fold, possessing a tetramerization domain, structure overview Methanothermobacter thermautotrophicus
tetramer 4 * 21000, static light scattering and SDS-PAGE Methanothermobacter thermautotrophicus

Synonyms

Synonyms Comment Organism
CobL
-
Methanothermobacter thermautotrophicus
precorrin-6y methyltransferase
-
Methanothermobacter thermautotrophicus
precorrin-8w decarboxylase
-
Methanothermobacter thermautotrophicus