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 (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 Maximum [Da] | Comment | Organism |
---|---|---|---|
21000 | - |
4 * 21000, static light scattering and SDS-PAGE | Methanothermobacter thermautotrophicus |
81000 | - |
static light scattering | Methanothermobacter thermautotrophicus |
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 | UniProt | Comment | Textmining |
---|---|---|---|
Methanothermobacter thermautotrophicus | - |
gene MT0146 or cbiT, fused to gene cbiE, resulting in cobL | - |
Purification (Comment) | Organism |
---|---|
recombinant enzyme CbiT from Escherichia coli strain B834 | Methanothermobacter thermautotrophicus |
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 | 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 | 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 | Comment | Organism |
---|---|---|
CobL | - |
Methanothermobacter thermautotrophicus |
precorrin-6y methyltransferase | - |
Methanothermobacter thermautotrophicus |
precorrin-8w decarboxylase | - |
Methanothermobacter thermautotrophicus |