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Literature summary for 2.1.1.202 extracted from

  • Awai, T.; Ochi, A.; Ihsanawati, A.; Sengoku, T.; Hirata, A.; Bessho, Y.; Yokoyama, S.; Hori, H.
    Substrate tRNA recognition mechanism of a multisite-specific tRNA methyltransferase, Aquifex aeolicus Trm1, based on the X-ray crystal structure (2011), J. Biol. Chem., 286, 35236-35246.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
gene MJ0026, expression in Escherichia coli strain BL21-CodonPlus(DE3)-RIL Methanocaldococcus jannaschii

Crystallization (Commentary)

Crystallization (Comment) Organism
Trm4 free and in complex with sinefungin, hanging-drop vapor diffusion, 0.001 ml of protein solution containing 10.8 mg/ml protein with 0.001 ml of crystallization solution containing 100 mM Tris-HCl, pH 7.4, 200 mM MgCl2, 25% PEG 3350, and 6% v/v isopropanol for MJ0026 alone, and 20 100 mM Tris-HCl, pH 7.4, 200 mM MgCl2, 25% PEG 3350, 3 mM sinefungin, and 10 mM cytidine for Mj0026 with sinefungin, cryoprotectant 20% glycerol and 20% PEG 400, respectively, X-ray diffraction structure determination and analysis at 1.27 A and 2.3 A resolutions, respectively Methanocaldococcus jannaschii

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
S-adenosyl-L-methionine + cytosine40 in tRNA precursor Methanocaldococcus jannaschii
-
S-adenosyl-L-homocysteine + 5-methylcytosine40 in tRNA precursor
-
?
S-adenosyl-L-methionine + cytosine48 in tRNA Methanocaldococcus jannaschii
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S-adenosyl-L-homocysteine + 5-methylcytosine48 in tRNA
-
?

Organism

Organism UniProt Comment Textmining
Methanocaldococcus jannaschii
-
gene MJ0026
-

Purification (Commentary)

Purification (Comment) Organism
recombinant MJ0026 from Escherichia coli strain BL21-CodonPlus(DE3)-RIL by heat treatment, anion exchange and hydroxyapatite chromatography, and dialysis Methanocaldococcus jannaschii

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information highly conserved residues form a large, positively charged surface, which seems to be suitable for tRNA binding. Substrate specificity and methylation sites on tRNA, overview Methanocaldococcus jannaschii ?
-
?
S-adenosyl-L-methionine + cytosine40 in tRNA precursor
-
Methanocaldococcus jannaschii S-adenosyl-L-homocysteine + 5-methylcytosine40 in tRNA precursor
-
?
S-adenosyl-L-methionine + cytosine48 in tRNA
-
Methanocaldococcus jannaschii S-adenosyl-L-homocysteine + 5-methylcytosine48 in tRNA
-
?

Synonyms

Synonyms Comment Organism
MJ0026
-
Methanocaldococcus jannaschii
RNA:m5C methyltransferase
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Methanocaldococcus jannaschii
TRM4
-
Methanocaldococcus jannaschii

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
70
-
assay at Methanocaldococcus jannaschii

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.7
-
assay at Methanocaldococcus jannaschii

Cofactor

Cofactor Comment Organism Structure
additional information the S-adenosyl-L-methionine analigue sinefungin is bound in a negatively charged pocket near helix alpha8, binding structure, detailed overview Methanocaldococcus jannaschii
S-adenosyl-L-methionine helix alpha8 can adopt two different conformations, thereby controlling the entry of S-adenosyl-L-methionine into the active site Methanocaldococcus jannaschii