Reference - Detail
|Author||Haque MR, Higashiura A, Nakagawa A, Hirowatari A, Furuya S, Yamamoto K.|
|Title||Molecular structure of a 5,10-methylenetetrahydrofolate dehydrogenase from the silkworm Bombyx mori.|
|Journal||FEBS Open Bio|
The enzyme 5,10-methylenetetrahydrofolate dehydrogenase (MTHFD) is essential for the production of certain amino acids (glycine, serine, and methionine) and nucleic acids (thymidylate and purine). Here, we identified a cDNA encoding this enzyme from the silkworm Bombyx mori. The recombinant B. mori MTHFD (bmMTHFD) expressed in Escherichia coli recognized 5,10-methylenetetrahydrofolate and 5,10-methenyltetrahydrofolate as substrate in the presence of NADP + as well as NAD +. The bmMTHFD structure was determined at a resolution of 1.75 Å by X-ray crystallography. Site-directed mutagenesis indicated that the amino acid residue Tyr49 contributed to its catalytic activity. Our findings provide insight into the mechanism underlying the activity of MTHFD from B. mori and potentially other insects and may therefore facilitate the development of inhibitors specific to MTHFD as insecticides.
|MeSH||Amino Acid Sequence Animals Bombyx / enzymology Bombyx / genetics* Bombyx / metabolism DNA, Complementary / chemistry DNA, Complementary / genetics DNA, Complementary / metabolism Escherichia coli / genetics Insect Proteins / chemistry Insect Proteins / genetics* Insect Proteins / metabolism Methylenetetrahydrofolate Dehydrogenase (NADP) / chemistry Methylenetetrahydrofolate Dehydrogenase (NADP) / genetics* Methylenetetrahydrofolate Dehydrogenase (NADP) / metabolism Molecular Structure Mutagenesis, Site-Directed Phylogeny Recombinant Proteins / chemistry Recombinant Proteins / genetics Recombinant Proteins / metabolism Sequence Alignment|