Biosynthesis of isoprenoids: a bifunctional IspDF enzyme from Campylobacter jejuni.

In the nonmevalonate pathway of isoprenoid biosynthesis, the conversion of 2C-methyl-d-erythritol 4-phosphate into its cyclic diphosphate proceeds via nucleotidyl intermediates and is catalyzed by the products of the ispD, ispE and ispF genes. An open reading frame of Campylobacter jejuni with similarity to the ispD and ispF genes of Escherichia coli was cloned into an expression vector directing the formation of a 42 kDa protein in a recombinant E. coli strain. The purified protein was shown to catalyze the transformation of 2C-methyl-D-erythritol 4-phosphate into 4-diphosphocytidyl-2C-methyl-D-erythritol and the conversion of 4-diphosphocytidyl-2C-methyl-D-erythritol 2-phosphate into 2C-methyl-D-erythritol 2,4-cyclodiphosphate at catalytic rates of 19 micro mol x mg(-1) x min(-1) and 7 micro mol x mg(-1) x min(-1), respectively. Both enzyme-catalyzed reactions require divalent metal ions. The C. jejuni enzyme does not catalyze the formation of 2C-methyl-D-erythritol 3,4-cyclophosphate from 4-diphosphocytidyl-2C-methyl-D-erythritol, a side reaction catalyzed in vitro by the IspF proteins of E. coli and Plasmodium falciparum. Comparative genomic analysis show that all sequenced alpha- and epsilon-proteobacteria have fused ispDF genes. These bifunctional proteins are potential drug targets in several human pathogens (e.g. Helicobacter pylori, C. jejuni and Treponema pallidum).

[1]  T. Bach Some new aspects of isoprenoid biosynthesis in plants—A review , 1995, Lipids.

[2]  A. Bacher,et al.  Biosynthesis of isoprenoids via the non-mevalonate pathway , 2004, Cellular and Molecular Life Sciences CMLS.

[3]  W. Hunter,et al.  Structure of a tetragonal crystal form of Escherichia coli 2-C-methyl-D-erythritol 4-phosphate cytidylyltransferase. , 2003, Acta crystallographica. Section D, Biological crystallography.

[4]  W. Eisenreich,et al.  The deoxyxylulose phosphate pathway of isoprenoid biosynthesis: Studies on the mechanisms of the reactions catalyzed by IspG and IspH protein , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[5]  W. Eisenreich,et al.  The deoxyxylulose phosphate pathway of isoprenoid biosynthesis. Discovery and function of the ispDEFGH genes and their cognate enzymes , 2003 .

[6]  Christian von Mering,et al.  STRING: a database of predicted functional associations between proteins , 2003, Nucleic Acids Res..

[7]  J. Wiesner,et al.  LytB protein catalyzes the terminal step of the 2‐C‐methyl‐D‐erythritol‐4‐phosphate pathway of isoprenoid biosynthesis , 2002, FEBS letters.

[8]  J. Wiesner,et al.  Functional characterization of GcpE, an essential enzyme of the non‐mevalonate pathway of isoprenoid biosynthesis , 2002, FEBS letters.

[9]  Jochen Wiesner,et al.  Fosmidomycin for malaria , 2002, The Lancet.

[10]  W. Eisenreich,et al.  Biosynthesis of terpenes: Studies on 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[11]  Simon F. Park The physiology of Campylobacter species and its relevance to their role as foodborne pathogens. , 2002, International journal of food microbiology.

[12]  Charles S Bond,et al.  Structure of 2C-methyl-d-erythritol 2,4- cyclodiphosphate synthase: An essential enzyme for isoprenoid biosynthesis and target for antimicrobial drug development , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[13]  M. Rohmer,et al.  Isoprenoid biosynthesis via the methylerythritol phosphate pathway. (E)-4-Hydroxy-3-methylbut-2-enyl diphosphate: chemical synthesis and formation from methylerythritol cyclodiphosphate by a cell-free system from Escherichia coli , 2002 .

[14]  B. Bourke Campylobacter infection: small bowel and colon. , 2002, Current opinion in gastroenterology.

[15]  S. Steinbacher,et al.  Structure of 2C-methyl-d-erythritol-2,4-cyclodiphosphate synthase involved in mevalonate-independent biosynthesis of isoprenoids. , 2002, Journal of molecular biology.

[16]  W. Eisenreich,et al.  Studies on the nonmevalonate terpene biosynthetic pathway: Metabolic role of IspH (LytB) protein , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[17]  W. Eisenreich,et al.  Studies on the nonmevalonate pathway to terpenes: The role of the GcpE (IspG) protein , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[18]  W. Eisenreich,et al.  Biosynthesis of terpenoids: efficient multistep biotransformation procedures affording isotope-labeled 2C-methyl-D-erythritol 4-phosphate using recombinant 2C-methyl-D-erythritol 4-phosphate synthase. , 2001, The Journal of organic chemistry.

[19]  C. Stancu,et al.  Statins: mechanism of action and effects , 2001, Journal of cellular and molecular medicine.

[20]  W. Eisenreich,et al.  Studies on the Non‐Mevalonate Pathway − Preparation and Properties of Isotope‐Labeled 2C‐Methyl‐D‐erythritol 2,4‐Cyclodiphosphate , 2001 .

[21]  J. Wiesner,et al.  LytB, a novel gene of the 2‐C‐methyl‐D‐erythritol 4‐phosphate pathway of isoprenoid biosynthesis in Escherichia coli , 2001, FEBS letters.

[22]  W. Eisenreich,et al.  Biosynthesis of terpenoids. 2C-Methyl-D-erythritol 2,4-cyclodiphosphate synthase (IspF) from Plasmodium falciparum. , 2001, European journal of biochemistry.

[23]  M. Rohmer,et al.  Identification of gcpE as a novel gene of the 2‐C‐methyl‐D‐erythritol 4‐phosphate pathway for isoprenoid biosynthesis in Escherichia coli , 2001, FEBS letters.

[24]  E. Gantt,et al.  Evidence of a Role for LytB in the Nonmevalonate Pathway of Isoprenoid Biosynthesis , 2000, Journal of bacteriology.

[25]  W. Eisenreich,et al.  Biosynthesis of terpenoids: 4-diphosphocytidyl-2C-methyl-D-erythritol synthase of Arabidopsis thaliana. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[26]  W. Eisenreich,et al.  Biosynthesis of terpenoids: YgbB protein converts 4-diphosphocytidyl-2C-methyl-D-erythritol 2-phosphate to 2C-methyl-D-erythritol 2,4-cyclodiphosphate. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[27]  W. Eisenreich,et al.  Biosynthesis of terpenoids: YchB protein of Escherichia coli phosphorylates the 2-hydroxy group of 4-diphosphocytidyl-2C-methyl-D-erythritol. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[28]  W. Eisenreich,et al.  Cytidine 5'-triphosphate-dependent biosynthesis of isoprenoids: YgbP protein of Escherichia coli catalyzes the formation of 4-diphosphocytidyl-2-C-methylerythritol. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[29]  H. Lichtenthaler,et al.  Inhibitors of the nonmevalonate pathway of isoprenoid biosynthesis as antimalarial drugs. , 1999, Science.

[30]  M. Rohmer 2.03 – A Mevalonate-independent Route to Isopentenyl Diphosphate , 1999 .

[31]  V. Rodwell,et al.  2.02 – Biosynthesis of Mevalonic Acid from Acetyl-CoA , 1999 .

[32]  M. Schwarz,et al.  2.14 – Ginkgolide Biosynthesis , 1999 .

[33]  W. Eisenreich,et al.  The deoxyxylulose phosphate pathway of terpenoid biosynthesis in plants and microorganisms. , 1998, Chemistry & biology.

[34]  S. Takahashi,et al.  A 1-deoxy-D-xylulose 5-phosphate reductoisomerase catalyzing the formation of 2-C-methyl-D-erythritol 4-phosphate in an alternative nonmevalonate pathway for terpenoid biosynthesis. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[35]  J. Sacchettini,et al.  Creating Isoprenoid Diversity , 1997, Science.

[36]  P. Kuzmič,et al.  Program DYNAFIT for the analysis of enzyme kinetic data: application to HIV proteinase. , 1996, Analytical biochemistry.

[37]  S. Shuman,et al.  Novel approach to molecular cloning and polynucleotide synthesis using vaccinia DNA topoisomerase. , 1994, The Journal of biological chemistry.

[38]  K. Bloch Sterol molecule: structure, biosynthesis, and function , 1992, Steroids.

[39]  F. Studier,et al.  Use of T7 RNA polymerase to direct expression of cloned genes. , 1990, Methods in enzymology.