title: Biosynthesis of terpenoids: 4-diphosphocytidyl-2-C-methyl-D-erythritol kinase from tomato.
abstract: The putative catalytic domain (residues 81-401) of a predicted tomato protein with similarity to 4-diphosphocytidyl-2-C-methyl-d-erythritol kinase of Escherichia coli was expressed in a recombinant E. coli strain. The protein was purified to homogeneity and was shown to catalyze the phosphorylation of the position 2 hydroxy group of 4-diphosphocytidyl-2-C-methyl-d-erythritol at a rate of 33 micromol small middle dotmg(-1) small middle dotmin(-1). The structure of the reaction product, 4-diphosphocytidyl-2-C-methyl-d-erythritol 2-phosphate, was established by NMR spectroscopy. Divalent metal ions, preferably Mg(2+), are required for activity. Neither the tomato enzyme nor the E. coli ortholog catalyzes the phosphorylation of isopentenyl monophosphate.
authors.affiliation: Lehrstuhl für Organische Chemie und Biochemie, Technische Universität München, Lichtenbergstrasse 4, D-85747 Garching, Germany.
authors.forename: F
authors.initials: F
authors.lastname: Rohdich
authors.forename: J
authors.initials: J
authors.lastname: Wungsintaweekul
authors.forename: H
authors.initials: H
authors.lastname: Luttgen
authors.forename: M
authors.initials: M
authors.lastname: Fischer
authors.forename: W
authors.initials: W
authors.lastname: Eisenreich
authors.forename: C A
authors.initials: CA
authors.lastname: Schuhr
authors.forename: M
authors.initials: M
authors.lastname: Fellermeier
authors.forename: N
authors.initials: N
authors.lastname: Schramek
authors.forename: M H
authors.initials: MH
authors.lastname: Zenk
authors.forename: A
authors.initials: A
authors.lastname: Bacher
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D018744:DNA, Plant; D004896:Erythritol / Q000031:analogs & derivatives* / Q000378:metabolism; D029968:Escherichia coli Proteins; D018551:Solanum lycopersicum / Q000201:enzymology* / Q000235:genetics; D008969:Molecular Sequence Data; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D011494:Protein Kinases / Q000378:metabolism; D013729:Terpenes / Q000378:metabolism
chemical_list: C400182:4-diphosphocytidyl-2-C-methylerythritol; D018744:DNA, Plant; D029968:Escherichia coli Proteins; D013729:Terpenes; D011494:Protein Kinases; D017853:Phosphotransferases (Alcohol Group Acceptor); C402428:isopentenyl monophosphate kinase; C404502:IspE protein, E coli; D004896:Erythritol