Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Taylor, A.B.; Meyer, B.; Leal, B.Z.; Koetter, P.; Schirf, V.; Demeler, B.; Hart, P.J.; Entian, K.D.; Woehnert, J.
    The crystal structure of Nep1 reveals an extended SPOUT-class methyltransferase fold and a pre-organized SAM-binding site (2008), Nucleic Acids Res., 36, 1542-1554.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.1.1.260 gene NEP1, DNA and amino acid sequence determination and analysis, expression in Escherichia coli strain BL21(DE3) Methanocaldococcus jannaschii

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.1.1.260 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

EC Number Inhibitors Comment Organism Structure
2.1.1.260 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/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.1.1.260 S-adenosyl-L-methionine + pseudouridine1191 in yeast 18S rRNA Methanocaldococcus jannaschii
-
 S-adenosyl-L-homocysteine + N1-methylpseudouridine1191 in yeast 18S rRNA
-
 ?
2.1.1.260 S-adenosyl-L-methionine + pseudouridine1191 in yeast 18S rRNA Methanocaldococcus jannaschii DSM 2661
-
 S-adenosyl-L-homocysteine + N1-methylpseudouridine1191 in yeast 18S rRNA
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.1.1.260 Methanocaldococcus jannaschii Q57977
-
-
2.1.1.260 Methanocaldococcus jannaschii DSM 2661 Q57977
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.1.1.260 recombinant Nep1p from Escherichia coli strain BL21(DE3) by ion exchange chromatography and gel filtration Methanocaldococcus jannaschii

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.1.1.260 S-adenosyl-L-methionine + pseudouridine1191 in yeast 18S rRNA
-
 Methanocaldococcus jannaschii S-adenosyl-L-homocysteine + N1-methylpseudouridine1191 in yeast 18S rRNA
-
 ?
2.1.1.260 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 ?
2.1.1.260 S-adenosyl-L-methionine + pseudouridine1191 in yeast 18S rRNA
-
 Methanocaldococcus jannaschii DSM 2661 S-adenosyl-L-homocysteine + N1-methylpseudouridine1191 in yeast 18S rRNA
-
 ?
2.1.1.260 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

EC Number Subunits Comment Organism
2.1.1.260 More Nep1 monomer and dimer structures, detailed overview Methanocaldococcus jannaschii

Synonyms

EC Number Synonyms Comment Organism
2.1.1.260 Nep1
-
 Methanocaldococcus jannaschii

Cofactor

EC Number Cofactor Comment Organism Structure
2.1.1.260 S-adenosyl-L-methionine
-
 Methanocaldococcus jannaschii

General Information

EC Number General Information Comment Organism
2.1.1.260 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
2.1.1.260 additional information active site structure and ligand binding, overview Methanocaldococcus jannaschii
2.1.1.260 physiological function Nep1 is a genuine rRNA methyltransferase and is essential for ribosome biogenesis Methanocaldococcus jannaschii