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Literature summary for 2.1.1.57 extracted from

  • Lockless, S.W.; Cheng, H.T.; Hodel, A.E.; Quiocho, F.A.; Gershon, P.D.
    Recognition of capped RNA substrates by VP39, the vaccinia virus-encoded mRNA cap-specific 2'-O-methyltransferase (1998), Biochemistry, 37, 8564-8574.
    View publication on PubMed
Search for more literature for 2.1.1.57

Cloned(Commentary)

Cloned (Comment) Organism
mutant enzymes are expressed in Escherichia coli and the tag is cleaved off Vaccinia virus

Protein Variants

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 [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

Organism UniProt Comment Textmining
Vaccinia virus
-
 VP39 protein
-

Purification (Commentary)

Purification (Comment) Organism
mutants recombinant from E. coli Vaccinia virus

Reaction

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
a

Substrates and Products (Substrate)

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

Synonyms Comment Organism
mRNA cap-specific 2'-O-methyltransferase i.e. VP39 Vaccinia virus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Vaccinia virus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.6
-
 assay at Vaccinia virus

pH Range

pH Minimum pH Maximum Comment Organism
6.5 8 m7G- and A-capped RNA substrates Vaccinia virus

pH Stability

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