Cloned (Comment) | Organism |
---|---|
gene NEP1, recombinaant expression of Nep1 | Methanocaldococcus jannaschii |
gene NEP1, recombinant expression of His-tagged Nep1 in Escherichia coli strain M15(pREP4) | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
D86G | the mutation causes the Bowen-Conradi syndrome | Homo sapiens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
S-adenosyl-L-methionine + pseudouridine1191 in yeast 18S rRNA | Homo sapiens | - |
S-adenosyl-L-homocysteine + N1-methylpseudouridine1191 in yeast 18S rRNA | - |
? | |
S-adenosyl-L-methionine + pseudouridine1191 in yeast 18S rRNA | Methanocaldococcus jannaschii | - |
S-adenosyl-L-homocysteine + N1-methylpseudouridine1191 in yeast 18S rRNA | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | - |
- |
- |
Methanocaldococcus jannaschii | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged Nep1 from Escherichia coli strain M15(pREP4) by nickel affinity chromatography | Homo sapiens |
recombinant Nep1 by cation exchange chromatography and gel filtration | Methanocaldococcus jannaschii |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | RNA-binding specificity by in vitro using fluorescence quenching assays and yeast three-hybrid screening. Identification of the binding site for methylation target RNAs by NMR-spectroscopy, overview | Homo sapiens | ? | - |
? | |
additional information | the Methanocaldococcus jannaschii Nep1 binds to the Saccharomyces cerevisiae Nep1 RNA consensus sequence 5'-UUCAAC-3' in the Methanocaldococcus jannaschii 16S rRNA. The region of Methanocaldococcus jannaschii 16S rRNA is equivalent to nt 1190-1195 in yeast 18S rRNA including the m1acp3-psi nucleotide at position 1191. And binding of MjNep1 to a 6mer RNA containing the consensus sequence 5'-UUCAAC-3'. Even a 5'-truncated RNA lacking the first uridine of the consensus sequence is bound with comparable affinity. RNA-binding specificity by in vitro using fluorescence quenching assays and yeast three-hybrid screening. Identification of the binding site for methylation target RNAs by NMR-spectroscopy, overview | Methanocaldococcus jannaschii | ? | - |
? | |
S-adenosyl-L-methionine + pseudouridine1191 in yeast 18S rRNA | - |
Homo sapiens | S-adenosyl-L-homocysteine + N1-methylpseudouridine1191 in yeast 18S rRNA | - |
? | |
S-adenosyl-L-methionine + pseudouridine1191 in yeast 18S rRNA | - |
Methanocaldococcus jannaschii | S-adenosyl-L-homocysteine + N1-methylpseudouridine1191 in yeast 18S rRNA | - |
? | |
S-adenosyl-L-methionine + pseudouridine1191 in yeast 18S rRNA | - |
Methanocaldococcus jannaschii | S-adenosyl-L-homocysteine + N1-methylpseudouridine1191 in yeast 18S rRNA | product identification by 1H-NMR-spectroscopy | ? | |
S-adenosyl-L-methionine + pseudouridine1191 in yeast 18S rRNA | RNA-oligonucleotides corresponding to nucleotides 1245-1255 of human 18S rRNA and containing pseudouridine (5'-GACWCAACACG-3') at position 1248. This position corresponds to nucleotide 914 of Methanocaldococcus jannaschii 16S rRNA and to nt 1191 of yeast 18S rRNA | Homo sapiens | S-adenosyl-L-homocysteine + N1-methylpseudouridine1191 in yeast 18S rRNA | product identification by MALDI-mass spectrometrical analysis | ? |
Subunits | Comment | Organism |
---|---|---|
dimer | three-dimensional structure of the Nep1 dimer, overview | Homo sapiens |
dimer | three-dimensional structure of the Nep1 dimer, overview | Methanocaldococcus jannaschii |
Synonyms | Comment | Organism |
---|---|---|
Emg1 | - |
Homo sapiens |
Emg1 | - |
Methanocaldococcus jannaschii |
Nep1 | - |
Homo sapiens |
Nep1 | - |
Methanocaldococcus jannaschii |
Nep1 methyltransferase | - |
Homo sapiens |
Nep1 methyltransferase | - |
Methanocaldococcus jannaschii |
pseudouridine-N1-specific methyltransferase | - |
Homo sapiens |
pseudouridine-N1-specific methyltransferase | - |
Methanocaldococcus jannaschii |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Homo sapiens |
65 | - |
assay at | Methanocaldococcus jannaschii |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Homo sapiens |
7.5 | - |
assay at | Methanocaldococcus jannaschii |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
S-adenosyl-L-methionine | - |
Homo sapiens | |
S-adenosyl-L-methionine | - |
Methanocaldococcus jannaschii |
General Information | Comment | Organism |
---|---|---|
evolution | Nep1 belongs to the SPOUT-class RNA methyltransferases, Nep1 subfamily | Homo sapiens |
evolution | Nep1 belongs to the SPOUT-class RNA methyltransferases, Nep1 subfamily | Methanocaldococcus jannaschii |
physiological function | MjNep1 binds its RNA target along an extended basic surface cleft at the dimer interface that involves both insertions to the SPOUT-class core fold in an orientation consistent with the proposed methylation of nt 914 in Methanocaldococcus jannaschii 16S rRNA | Methanocaldococcus jannaschii |