Cloned (Comment) | Organism |
---|---|
mutant enzymes are expressed in Escherichia coli and the tag is cleaved off | Vaccinia virus |
Protein Variants | Comment | Organism |
---|---|---|
D182A | m7G binding pocket mutant, mutagenesis, C-terminal truncated by 26 amino acids, lacks specific m7G-contact side chains and shows reduced activity, but remains cap-dependent | Vaccinia virus |
D182A/E233A | m7G binding pocket double mutant, mutagenesis, N-terminal glutathione-S-transferase-tagged, C-terminal truncated by 26 amino acids, lacks specific m7G-contact side chains and shows reduced activity, but remains cap-dependent | Vaccinia virus |
E233A | m7G binding pocket mutant, mutagenesis, N-terminal glutathione-S-transferase-tagged, C-terminal truncated by 26 amino acids, lacks specific m7G-contact side chains and shows reduced activity, but remains cap-dependent | Vaccinia virus |
F180A | m7G binding pocket mutant, mutagenesis, C-terminal truncated by 26 amino acids, lacks specific m7G-contact side chains and shows reduced activity, but remains cap-dependent | Vaccinia virus |
Y22A | m7G binding pocket mutant, mutagenesis, N-terminal glutathione-S-transferase-tagged, C-terminal truncated by 26 amino acids, lacks specific m7G-contact side chains and shows reduced activity, but remains cap-dependent | Vaccinia virus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | - |
Vaccinia virus | |
0.000005 | 0.000015 | RNA chain length 20-50 nt | - |
Vaccinia virus | |
0.00016 | 0.00023 | RNA chain length 2-6 nt | - |
Vaccinia virus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Vaccinia virus | - |
VP39 protein | - |
Purification (Comment) | Organism |
---|---|
mutants recombinant from E. coli | Vaccinia virus |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
S-adenosyl-L-methionine + a 5'-(N7-methyl 5'-triphosphoguanosine)-(ribonucleotide)-[mRNA] = S-adenosyl-L-homocysteine + a 5'-(N7-methyl 5'-triphosphoguanosine)-(2'-O-methyl-ribonucleotide)-[mRNA] | binding sites of the enzyme for the terminal cap nucleotide, m7G, the cap-proximal RNA, and for RNA, immediately downstream, are characterized | Vaccinia virus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
S-adenosyl-L-methionine + ApppGR-RNA | adenine-capped RNA, wild-type and mutants, indicating the redundancy of m7G-contact residues able to confer cap-type specificity | Vaccinia virus | ? | - |
? | |
S-adenosyl-L-methionine + ApppGR-RNA | synthetic substrate with unusual cap structure, 20% activity compared to m7G-terminated substrates | Vaccinia virus | ? | - |
? | |
S-adenosyl-L-methionine + m7G(5')pppR-RNA | adenine-capped RNA, wild-type and mutants, indicating the redundancy of m7G-contact residues able to confer cap-type specificity | Vaccinia virus | S-adenosyl-L-homocysteine + m7G(5')pppRm-RNA | - |
? | |
S-adenosyl-L-methionine + m7GpppGp | substrate of minimal chain length for the cap-specific enzyme, m7GpppG is no substrate | Vaccinia virus | S-adenosyl-L-homocysteine + m7GpppGmp | - |
? | |
S-adenosyl-L-methionine + m7GpppGpN | - |
Vaccinia virus | S-adenosyl-L-homocysteine + m7GpppGmpN | - |
? | |
S-adenosyl-L-methionine + m7GpppGpNp | - |
Vaccinia virus | ? | - |
? | |
S-adenosyl-L-methionine + m7GpppGpUbiotin-p | - |
Vaccinia virus | ? | - |
? | |
S-adenosyl-L-methionine + RNA chain length 2-6 nt | - |
Vaccinia virus | ? | - |
r | |
S-adenosyl-L-methionine + RNA chain length 20-50 nt | - |
Vaccinia virus | ? | - |
r |
Synonyms | Comment | Organism |
---|---|---|
mRNA cap-specific 2'-O-methyltransferase | i.e. VP39 | Vaccinia virus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Vaccinia virus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.6 | - |
assay at | Vaccinia virus |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6.5 | 8 | m7G- and A-capped RNA substrates | Vaccinia virus |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
additional information | - |
binding site of the enzyme for RNA, immediately downstream, is pH-sensitive, above pH 7.5: dramatic increase of Km | Vaccinia virus |