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Deuterium-labelled isotopomers of 2-C-methyl-D-erythritol as tools for the elucidation of the 2-C-methyl-D-erythritol 4-phosphate pathway for isoprenoid biosynthesis.
Biochem J. 2000 Mar 15; 346 Pt 3:737-42.BJ

Abstract

Escherichia coli synthesizes its isoprenoids via the mevalonate-independent 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway. The MC4100dxs::CAT strain, defective in deoxyxylulose-5-phosphate synthase, which is the first enzyme in this metabolic route, exclusively synthesizes its isoprenoids from exogenous 2-C-methyl-D-erythritol (ME) added to the culture medium. The fate of the hydrogen atoms in the MEP pathway was followed by the incorporation of [1,1-(2)H(2)]ME and [3,5,5,5-(2)H(4)]ME. The two C-1 hydrogen atoms of ME were found without any loss in the prenyl chain of menaquinone and/or ubiquinone on the carbon atoms derived from C-4 of isopentenyl diphosphate (IPP) and on the E-methyl group of dimethylallyl diphosphate (DMAPP), the C-5 hydrogen atoms on the methyl groups derived from IPP C-5 methyl group and the Z-methyl group of DMAPP. This showed that no changes in the oxidation state of these carbon atoms occurred in the reaction sequence between MEP and IPP. Furthermore, no deuterium scrambling was observed between the carbon atoms derived from C-4 and C-5 of IPP or DMAPP, suggesting a completely stereoselective IPP isomerase or no significant activity of this enzyme. The C-3 deuterium atom of [3,5,5,5-(2)H(4)]ME was preserved only in the DMAPP starter unit and was completely missing from all those derived from IPP. This finding, aided by the non-essential role of the IPP isomerase gene, suggests the presence in E. coli of two different routes towards IPP and DMAPP, starting from a common intermediate derived from MEP.

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Authors+Show Affiliations

Charon L
Université Louis Pasteur/CNRS, Institut Le Bel, 4 rue Blaise Pascal, 67070 Strasbourg Cedex, France.
Hoeffler JF
No affiliation info available
Pale-Grosdemange C
No affiliation info available
Lois LM
No affiliation info available
Campos N
No affiliation info available
Boronat A
No affiliation info available
Rohmer M
No affiliation info available

MeSH

Base SequenceDNA PrimersDeuteriumErythritolEscherichia coliGenes, BacterialMagnetic Resonance SpectroscopyMass SpectrometryMutagenesis, Site-DirectedSugar PhosphatesTerpenes

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

10698701

Citation

Charon, L, et al. "Deuterium-labelled Isotopomers of 2-C-methyl-D-erythritol as Tools for the Elucidation of the 2-C-methyl-D-erythritol 4-phosphate Pathway for Isoprenoid Biosynthesis." The Biochemical Journal, vol. 346 Pt 3, 2000, pp. 737-42.
Charon L, Hoeffler JF, Pale-Grosdemange C, et al. Deuterium-labelled isotopomers of 2-C-methyl-D-erythritol as tools for the elucidation of the 2-C-methyl-D-erythritol 4-phosphate pathway for isoprenoid biosynthesis. Biochem J. 2000;346 Pt 3:737-42.
Charon, L., Hoeffler, J. F., Pale-Grosdemange, C., Lois, L. M., Campos, N., Boronat, A., & Rohmer, M. (2000). Deuterium-labelled isotopomers of 2-C-methyl-D-erythritol as tools for the elucidation of the 2-C-methyl-D-erythritol 4-phosphate pathway for isoprenoid biosynthesis. The Biochemical Journal, 346 Pt 3, 737-42.
Charon L, et al. Deuterium-labelled Isotopomers of 2-C-methyl-D-erythritol as Tools for the Elucidation of the 2-C-methyl-D-erythritol 4-phosphate Pathway for Isoprenoid Biosynthesis. Biochem J. 2000 Mar 15;346 Pt 3:737-42. PubMed PMID: 10698701.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Deuterium-labelled isotopomers of 2-C-methyl-D-erythritol as tools for the elucidation of the 2-C-methyl-D-erythritol 4-phosphate pathway for isoprenoid biosynthesis. AU - Charon,L, AU - Hoeffler,J F, AU - Pale-Grosdemange,C, AU - Lois,L M, AU - Campos,N, AU - Boronat,A, AU - Rohmer,M, PY - 2000/3/4/pubmed PY - 2000/5/8/medline PY - 2000/3/4/entrez SP - 737 EP - 42 JF - The Biochemical journal JO - Biochem J VL - 346 Pt 3 N2 - Escherichia coli synthesizes its isoprenoids via the mevalonate-independent 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway. The MC4100dxs::CAT strain, defective in deoxyxylulose-5-phosphate synthase, which is the first enzyme in this metabolic route, exclusively synthesizes its isoprenoids from exogenous 2-C-methyl-D-erythritol (ME) added to the culture medium. The fate of the hydrogen atoms in the MEP pathway was followed by the incorporation of [1,1-(2)H(2)]ME and [3,5,5,5-(2)H(4)]ME. The two C-1 hydrogen atoms of ME were found without any loss in the prenyl chain of menaquinone and/or ubiquinone on the carbon atoms derived from C-4 of isopentenyl diphosphate (IPP) and on the E-methyl group of dimethylallyl diphosphate (DMAPP), the C-5 hydrogen atoms on the methyl groups derived from IPP C-5 methyl group and the Z-methyl group of DMAPP. This showed that no changes in the oxidation state of these carbon atoms occurred in the reaction sequence between MEP and IPP. Furthermore, no deuterium scrambling was observed between the carbon atoms derived from C-4 and C-5 of IPP or DMAPP, suggesting a completely stereoselective IPP isomerase or no significant activity of this enzyme. The C-3 deuterium atom of [3,5,5,5-(2)H(4)]ME was preserved only in the DMAPP starter unit and was completely missing from all those derived from IPP. This finding, aided by the non-essential role of the IPP isomerase gene, suggests the presence in E. coli of two different routes towards IPP and DMAPP, starting from a common intermediate derived from MEP. SN - 0264-6021 UR - https://www.unboundmedicine.com/medline/citation/10698701/Deuterium_labelled_isotopomers_of_2_C_methyl_D_erythritol_as_tools_for_the_elucidation_of_the_2_C_methyl_D_erythritol_4_phosphate_pathway_for_isoprenoid_biosynthesis_ L2 - https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/10698701/ DB - PRIME DP - Unbound Medicine ER -
Grapherence [↓15 ↑17]

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