Cloned (Comment) | Organism |
---|---|
gene rsmD, complementation of rsmD-deleted Escherichia coli KL16DELTArsmD cells, expression of Rv2966c with His10-Smt3 fusion at the N-terminus from plasmid pMTase1 in Escherichia coli strain BL21 (DE3) | Mycobacterium tuberculosis |
Crystallization (Comment) | Organism |
---|---|
purified recombinant Rv2966c, hanging drop vapour diffusion method, mixing of 0.004 ml of 17 mg/ml Rv2966c in 10 mM potassium phosphate buffer, pH 7.4, 5 mM EDTA, 10% v/v glycerol, 0.1% v/v 2-mercaptoethanol, 50 mM NaCl, 0.1 mM S-adenosyl-L-methionine, with 0.002 ml of reservoir solution containing 100 mM Tris-HCl, pH 7.4, 3.0 M potassium acetate at 24 °C, 2-3 days, X-ray diffraction structure determination and analysis at 1.9 A resolution, molecular replacement | Mycobacterium tuberculosis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mycobacterium tuberculosis | P95128 | gene rv2966c | - |
Purification (Comment) | Organism |
---|---|
recombinant Rv2966c with His10-Smt3 fusion at the N-terminus from Escherichia coli strain BL21 (DE3) by nickel affinity and anion exchange chromatography | Mycobacterium tuberculosis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | the N-terminal domain of Rv2966c is important for binding not only RNA but also DNA substrates, binding of protein to DNA is much stronger than it is to RNA | Mycobacterium tuberculosis | ? | - |
? | |
S-adenosyl-L-methionine + guanine966 in 16S rRNA | recombinant Rv2966c can methylate G966 of 16S rRNA in vitro | Mycobacterium tuberculosis | S-adenosyl-L-homocysteine + N2-methylguanine966 in 16S rRNA | - |
? | |
S-adenosyl-L-methionine + guanine966 in 30S rRNA | recombinant Rv2966c can use 30S ribosomes purified from rsmD-deleted Escherichia coli as substrate | Mycobacterium tuberculosis | S-adenosyl-L-homocysteine + N2-methylguanine966 in 30S rRNA | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | protein Rv2966c shows a two-domain structure with a short hairpin domain at the N-terminus and a C-terminal domain with the S-adenosylmethionine-methyltransferase-fold. The N-terminal hairpin is a minimalist functional domain that helps Rv2966c in target recognition. In contrast to the variable substrate binding domain, the C-terminal domain is a highly conserved structure consisting of the classical S-adenosyl-L-methionine-methyltransferase-fold found in several S-adenosyl-L-methionine-dependent methyltransferases. It consists of a central eight-stranded beta-sheet flanked by alpha-helices on both sides, the first five strands of the beta-sheet are parallel to each other and encompass the S-adenosyl-L-methionine binding site toward their C terminus end, whereas the remaining strands of the beta-sheet are antiparallel. The N-terminus of Rv2966c is the target recognition region that helps in binding to nucleic acids | Mycobacterium tuberculosis |
Synonyms | Comment | Organism |
---|---|---|
m2G966 methyltransferase | - |
Mycobacterium tuberculosis |
RsmD-like methyltransferase | - |
Mycobacterium tuberculosis |
Rv2966c | - |
Mycobacterium tuberculosis |
Rv2966c protein | - |
Mycobacterium tuberculosis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Mycobacterium tuberculosis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Mycobacterium tuberculosis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
S-adenosyl-L-methionine | binding structure, overview | Mycobacterium tuberculosis |
General Information | Comment | Organism |
---|---|---|
evolution | structural similarities with other methyltransferases are limited to the core S-adenosyl-L-methionine domain but not the target recognition domain | Mycobacterium tuberculosis |
malfunction | truncated constructs of Rv2966c lacking the N-terminal 18- or 23-amino acid residues and the mutant protein 3RM do not show any significant binding to either DNA or RNA nor any m2G966 methyltransferase activity | Mycobacterium tuberculosis |
additional information | the N-terminal hairpin is a minimalist functional domain that helps Rv2966c in target recognition | Mycobacterium tuberculosis |