Studies on the nonmevalonate pathway to terpenes: the role of the GcpE (IspG) protein.
Recombinant Escherichia coli cells engineered for the expression of the xylB gene in conjunction with genes of the nonmevalonate pathway were supplied with (13)C-labeled 1-deoxy-D-xylulose. Cell extracts were analyzed directly by NMR spectroscopy. (13)C-labeled 2C-methyl-D-erythritol 2,4-cyclodiphosphate was detected at high levels in cells expressing xylB, ispC, ispD, ispE, and ispF. The additional expression of the gcpE gene afforded 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate as an intermediate of the nonmevalonate pathway. Hypothetical mechanisms involving conserved cysteine residues are proposed for the enzymatic conversion of 2C-methyl-D-erythritol 2,4-cyclodiphosphate into 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate catalyzed by the GcpE protein.
Hecht
S
S
Lehrstuhl für Organische Chemie und Biochemie, Technische Universität München, Lichtenbergstrasse 4, D-85747 Garching, Germany.
Eisenreich
W
W
Adam
P
P
Amslinger
S
S
Kis
K
K
Bacher
A
A
Arigoni
D
D
Rohdich
F
F
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000502:physiology*; D001483:Base Sequence; D017931:DNA Primers; D004798:Enzymes; D008798:Mevalonic Acid / Q000378:metabolism*; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D017386:Sequence Homology, Amino Acid; D013729:Terpenes / Q000378:metabolism*
D001426:Bacterial Proteins; D017931:DNA Primers; D004798:Enzymes; D013729:Terpenes; C421242:hydroxymethylbutenyl 4-diphosphate synthase; D008798:Mevalonic Acid