Biosynthesis of terpenoids: 4-diphosphocytidyl-2-C-methyl-D-erythritol kinase from tomato.
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.
Rohdich
F
F
Lehrstuhl für Organische Chemie und Biochemie, Technische Universität München, Lichtenbergstrasse 4, D-85747 Garching, Germany.
Wungsintaweekul
J
J
Luttgen
H
H
Fischer
M
M
Eisenreich
W
W
Schuhr
C A
CA
Fellermeier
M
M
Schramek
N
N
Zenk
M H
MH
Bacher
A
A
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
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