Cloned (Comment) | Organism |
---|---|
gene NEP1, DNA and amino acid sequence determination and analysis, expression in Escherichia coli strain BL21(DE3) | Methanocaldococcus jannaschii |
Crystallization (Comment) | Organism |
---|---|
Nep1 in its free form and bound to S-adenosylhomocysteine or the antibiotic and general methyltransferase inhibitor sinefungin, hanging drop vapour diffusion method, 10-15 mg/ml protein in 100 mM BisTris-propane buffer, pH 9.5, 6-10% PEG 400, 30-48% glycerol, and 0-400 mM trimethylamine-N-oxide, at 4°C, crystals of the selenomethionine containing protein in the presence or absence of a 3fold excess of S-adenosyl-L-homocysteine are grown under similar conditions at 16°C, X-ray diffraction structure determination and analysis at 2.2 A, 2.15 A, and 2.25 A resolution, respectively | Methanocaldococcus jannaschii |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
sinefungin | sinefungin binds to Nep1 at a preformed binding site that is topologically equivalent to the cofactor binding site in other SPOUT-class methyltransferases | Methanocaldococcus jannaschii |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
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 DSM 2661 | - |
S-adenosyl-L-homocysteine + N1-methylpseudouridine1191 in yeast 18S rRNA | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Methanocaldococcus jannaschii | Q57977 | - |
- |
Methanocaldococcus jannaschii DSM 2661 | Q57977 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant Nep1p from Escherichia coli strain BL21(DE3) by ion exchange chromatography and gel filtration | Methanocaldococcus jannaschii |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
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 | S-adenosyl-L-methionine binding site structure, overview | Methanocaldococcus jannaschii | S-adenosyl-L-homocysteine + N1-methylpseudouridine1191 in yeast 18S rRNA | S-adenosyl-L-homocysteine binds to Nep1 at a preformed binding site that is topologically equivalent to the cofactor binding site in other SPOUT-class methyltransferases, intermolecular interactions, overview | ? | |
S-adenosyl-L-methionine + pseudouridine1191 in yeast 18S rRNA | - |
Methanocaldococcus jannaschii DSM 2661 | S-adenosyl-L-homocysteine + N1-methylpseudouridine1191 in yeast 18S rRNA | - |
? | |
S-adenosyl-L-methionine + pseudouridine1191 in yeast 18S rRNA | S-adenosyl-L-methionine binding site structure, overview | Methanocaldococcus jannaschii DSM 2661 | S-adenosyl-L-homocysteine + N1-methylpseudouridine1191 in yeast 18S rRNA | S-adenosyl-L-homocysteine binds to Nep1 at a preformed binding site that is topologically equivalent to the cofactor binding site in other SPOUT-class methyltransferases, intermolecular interactions, overview | ? |
Subunits | Comment | Organism |
---|---|---|
More | Nep1 monomer and dimer structures, detailed overview | Methanocaldococcus jannaschii |
Synonyms | Comment | Organism |
---|---|---|
Nep1 | - |
Methanocaldococcus jannaschii |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
S-adenosyl-L-methionine | - |
Methanocaldococcus jannaschii |
General Information | Comment | Organism |
---|---|---|
malfunction | addition of SAM to the medium restores growth at elevated temperatures in yeast with temperature sensitive mutants of the yeast Nep1 protein | Methanocaldococcus jannaschii |
additional information | active site structure and ligand binding, overview | Methanocaldococcus jannaschii |
physiological function | Nep1 is a genuine rRNA methyltransferase and is essential for ribosome biogenesis | Methanocaldococcus jannaschii |