Cloned (Comment) | Organism |
---|---|
expression of His-tagged full length enzyme, residues 211-597, comprising the catalytical domain, and residues 1-210 in Escherichia coli BL21(DE3) | Mus musculus |
expression of wild-type full length enzyme and catalytic domain in deficient Saccharomyces cerevisiae strain YBS2 as His-tagged protein, functional complementation by both of them | Mus musculus |
mRNA capping enzyme, DNA sequence determination and analysis | Mus musculus |
Protein Variants | Comment | Organism |
---|---|---|
K294A | inactive, no functional complementation of the deficient Saccharomyces cerevisiae mutant | Mus musculus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | optimal at 5-10 mM | Mus musculus | |
Mg2+ | enzyme-GMP intermediate formation is strictly dependent on divalent cations, satisfied by Mn2+ or Mg2+, Mn2+ is slightly preferred | Mus musculus | |
Mn2+ | enzyme-GMP intermediate formation is strictly dependent on divalent cations, satisfied by Mn2+ or Mg2+, Mn2+ is slightly preferred | Mus musculus | |
Mn2+ | optimal at 1-2 mM | Mus musculus | |
additional information | requirement for divalent cations | Mus musculus |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
28000 | - |
catalytically active residues 211-597, i.e. RNA guanylyltransferase domain, glycerol gradient sedimentation | Mus musculus |
46000 | - |
residues 1-210, i.e. RNA triphosphatase domain, glycerol gradient sedimentation | Mus musculus |
68000 | - |
mRNA capping enzyme, glycerol density gradient sedimentation | Mus musculus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
GTP + pp(5')RNA | Mus musculus | mouse guanylyltransferase domain can substitute the deficient enzyme in Saccharomyces cerevisiae mutant in vivo | G(5')ppp(5')RNA + diphosphate | guanosine residue linked 5' through three phosphates to the 5' position of the terminal residue | ? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mus musculus | O55236 | mRNA capping enzyme | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged full length enzyme as well as the recombinant His-tagged N-terminal and C-terminal domains from Escherichia coli BL21(DE3) | Mus musculus |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
GTP + (5')ppPur-mRNA = diphosphate + G(5')pppPur-mRNA | guanylyltransferase domain: residues 211-597 | Mus musculus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
GTP + pp(5')RNA | the enzymes isolated guanylyltransferase domian, residues 211-597, is catalytically active in vitro | Mus musculus | G(5')ppp(5')RNA + diphosphate | guanosine residue linked 5' through three phosphates to the 5' position of the terminal residue | ? | |
GTP + pp(5')RNA | formation of a covalent intermediate enzyme-GMP | Mus musculus | G(5')ppp(5')RNA + diphosphate | guanosine residue linked 5' through three phosphates to the 5' position of the terminal residue | ? | |
GTP + pp(5')RNA | mouse guanylyltransferase domain can substitute the deficient enzyme in Saccharomyces cerevisiae mutant in vivo | Mus musculus | G(5')ppp(5')RNA + diphosphate | guanosine residue linked 5' through three phosphates to the 5' position of the terminal residue | ? |
Subunits | Comment | Organism |
---|---|---|
monomer | 1 * 46000, catalytically active residues 211-597, i.e. guanylyltransferase domain, + 1 * 28000, residues 1-210, i.e. RNA triphosphatase domain, SDS-PAGE | Mus musculus |
Synonyms | Comment | Organism |
---|---|---|
More | enzyme is part of the bifunctional capping enzyme, which consists of autonomous and non-overlapping RNA 5'-triphosphatase and a RNA guanylyltransferase domain and activity | Mus musculus |