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Literature summary 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)

EC Number Cloned (Comment) Organism
2.1.1.132 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)

EC Number Crystallization (Comment) Organism
2.1.1.132 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
2.1.1.196 homotetrameric apo form of CbiT crystallized in several space groups, to about 2.5 A resolution, and in complex with S-adenosyl-L-homocysteine, to 1.9 A resolution. The protein shows structural similarity to Rossmann-like S-adenosyl-methionine-dependent methyltransferases, and the cocrystal structure shows that it binds S-adenosyl-methionine in standard geometry near a binding pocket that can accommodate a precorrin substrate. CbiT probably functions as a precorrin methyltransferase Methanothermobacter thermautotrophicus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.1.1.132 21000
-
 4 * 21000, static light scattering and SDS-PAGE Methanothermobacter thermautotrophicus
2.1.1.132 81000
-
 static light scattering Methanothermobacter thermautotrophicus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.1.1.132 additional information Methanothermobacter thermautotrophicus evolutionary origin of the cbiT ?
-
 ?
2.1.1.132 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

EC Number Organism UniProt Comment Textmining
2.1.1.132 Methanothermobacter thermautotrophicus
-
 gene MT0146 or cbiT, fused to gene cbiE, resulting in cobL
-
2.1.1.196 Methanothermobacter thermautotrophicus O26249
-
-

Purification (Commentary)

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

Reaction

EC Number Reaction Comment Organism Reaction ID
2.1.1.132 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
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.1.1.132 additional information evolutionary origin of the cbiT Methanothermobacter thermautotrophicus ?
-
 ?
2.1.1.132 additional information S-adenosyl-L-methionine binding structure and mechanism of CbiT Methanothermobacter thermautotrophicus ?
-
 ?
2.1.1.132 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
-
 ?
2.1.1.132 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

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

Synonyms

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